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Seed dispersal by vertebrates is fundamental for the persistence of plant species, forming networks of interactions that are often nested and modular. Networks involving angiosperms and frugivorous birds are relatively well-studied in the Neotropical region, but there are no previous studies of networks involving waterbirds. Here, we describe the structure of a Neotropical waterfowl seed-dispersal network and identify the species that have an important role for the network structure. We used information on 40 plant taxa found in fecal samples of five common waterfowl species to calculate the nestedness (NODF), weighted nestedness (WNODF), modularity, and weighted modularity of the network. We found that the network was nested, with yellow-billed teal showing the highest contribution both to nestedness and weighted nestedness. Twenty-four plant species contributed positively to weighted nestedness, with Salzmann's mille graines presenting the highest influence both to nestedness and weighted nestedness. The network was modular, but the weighted modularity was not significant. These results need to be considered with caution due to incomplete interaction sampling for two species. Ringed teal, Brazilian teal, and yellow-billed teal were considered hub modular species. Among plants, beak sedges and water snowflake were considered modular hub species, while Salzmann's mille graines and spikerush were network connectors. The structure of this Neotropical waterbird seed-dispersal network differed from the only previous waterfowl network study, from Europe, which found similar level of nestedness but no significant modularity. We include several possible explanations for this discrepancy and identified priorities for future research into waterbird–plant interaction networks. GGS received a postdoctoral fellowship from CNPq (Grant No. 150887/2022-1). MAP (Grant No. 304742/2019-8), LB (311409/2018-0) and LM thank the Brazilian Council for Development of Science and Technology (CNPq) for Research Productivity Grants. AJG was supported by Spanish National Plan projects CGL2016-76067-P (AEI/FEDER, EU) and PID2020-112774GB-I00/AEI/10.13039/501100011033. ESG received the grant RYC2019-027216-I funded by MCIN/AEI/10.13039/501100011033 and by ESF Investing in your future. This research was supported by funds from CNPq (Grant No. 52370695-2).

Understanding the factors that allow multiple species to coexist and share resources is an outstanding question in community ecology. Animals that share resources tend to use different strategies to decrease potential competition, through morphological adaptations, establishment of hierarchies, behavioral adaptations or spatial or temporal segregation. The main objective of this study was to infer interspecific processes of competition and facilitation through the study of species co-occurrence patterns in a vertebrate scavenger guild in de Brazilian cerrado. We analyzed patterns of spatial and temporal co-occurrence between species pairs, both qualitatively and quantitatively, and determined the activity patterns of the different scavenger species. For this purpose, we placed and monitored 11 large (i.e. goat) and 45 small (i.e. chicken) carcasses by camera-trapping, obtaining a total of 27 448 images. Our results show complex competitive and facilitative relationships among scavenging species in the Brazilian cerrado that are influenced by carcass size and change depending on the spatial and temporal scale at which they are analyzed. The scavenger assemblages that consumed large and small carcasses were different, evidencing resource partitioning between obligate and facultative scavengers. Furthermore, as an alternative to reduce competition levels, most species showed differences in their scavenging patterns, in addition to a strong temporal segregation during carcass consumption. Regarding New World vultures, our results suggest a strong interference competition between species with clear differences in their ecological traits (e.g. size, social behavior). However, we also found evidence of facilitation processes between vulture species in the location and access to the interior of the carcasses. Our findings highlight the role of obligate scavengers both in competition and facilitation processes in this vertebrate scavenger community. Future research should focus on investigating which species play the most important role in the structure and dynamics of this community, also considering intraspecific and behavioral patterns. LNA, ZMR and ESG were supported by the Generalitat Valenciana and the European Social Fund (ACIF/2019/056, APOSTD/2019/016, SEJI/2018/024, respectively), and JASZ by funds from the Spanish Ministry of Science, Innovation and Universities and the European Regional Development Fund (RTI2018-099609-B-C21). ESG received the grant RYC2019-027216-I funded by MCIN/AEI/10.13039/501100011033 and by ESF Investing in your future. ZMR was also supported by a postdoctoral contract funded by the Junta de Andalucía (POSTDOC_21_00353).

Background The long-term outcome of patients after antitumour necrosis factor alpha (anti-TNF) discontinuation is not well known. Aims To assess the risk of relapse in the long-term after anti-TNF discontinuation. Methods This was an extension of the evolution after anti-TNF discontinuation in patients with inflammatory bowel disease (EVODIS) study (Crohn's disease or ulcerative colitis patients treated with anti-TNFs in whom these drugs were withdrawn after achieving clinical remission) based in the same cohort of patients whose outcome was updated. Clinical remission was defined as a Harvey-Bradshaw index ≤4 points in Crohn's disease, a partial Mayo score ≤2 in ulcerative colitis and the absence of fistula drainage despite gentle finger compression in perianal disease. Results This was an observational, retrospective, multicenter study. A total of 1055 patients were included. The median follow-up time was 34 months. The incidence rate of relapse was 12% per patient-year (95% confidence interval [CI] = 11-14). The cumulative incidence of relapse was 50% (95% CI = 47-53): 19% at one year, 31% at 2 years, 38% at 3 years, 44% at 4 years and 48% at 5 years of follow-up. Of the 60% patients retreated with the same anti-TNF after relapse, 73% regained remission. Of the 75 patients who did not respond, 48% achieved remission with other therapies. Of the 190 patients who started other therapies after relapse, 62% achieved remission with the new treatment. Conclusions A significant proportion of patients who discontinued the anti-TNF remained in remission. In case of relapse, retreatment with the same anti-TNF was usually effective. Approximately half of the patients who did not respond after retreatment achieved remission with other therapies.

Rural abandonment and subsequent vegetation regeneration (‘passive rewilding') are expected to increase worldwide, producing cascades of dynamic socioeconomic, landscape and biological changes. Although landscape characteristics strongly influence the structure and functioning of scavengers, little is known about the ecological consequences of passive rewilding due to woody encroachment (i.e.‘landscape closure') on scavenging assemblages. We investigated differences in ‘scavenger assemblage composition' (species richness and abundances) and ‘scavenging efficiency' (scavenging frequency, detection and consumption times and consumption rates) in a mountain agroecosystem (Pyrenees) undergoing passive rewilding. We monitored 178 carcasses in three landscapes: ‘open', ‘shrubland' and ‘forest', and evaluated the effects of landscape type on ‘scavenger assemblage composition' and ‘scavenging efficiency' at the community and species levels, while accounting for the influences of carcass size, type and placement time. We also examined whether the locally most abundant and efficient scavenger (i.e. the griffon vulture Gyps fulvus) affects scavenging patterns. We found that landscape type was the main factor governing scavenging dynamics. Overall and average scavenger richness were similar in open and shrubland landscapes, while forests contained the lowest number of scavengers, mainly comprising mammals. Unlike mammals, avian scavenging frequency decreased as vegetation cover increased, especially for obligate scavengers (i.e. vultures). Scavenger abundances were highest in open landscapes, and carcasses were detected and consumed more rapidly in these landscapes. Carcass size did not influence detection and consumption times, although it did affect average scavenger richness, abundances and consumption rates. Consumption rates were higher in open landscapes and were strongly associated with the presence of griffon vultures. Interestingly, we found that griffon vultures influenced scavenging dynamics via facilitation processes. However, woody encroachment could reduce the scavenging role of this species, while favoring mammalian facultative scavengers. Finally, our findings highlight the pivotal role of griffon vultures, mediated by landscape characteristics, in reducing carcass persistence., POV and AM were funded within the framework of the project RTI2018- 099609-B-C22, from the I + D + I National Plan funded by the Ministry of Science, Innovation and Universities. ESG was funded by a Ramón y Cajal fellow (RYC-2019-027216-I).

The striatum is the largest entrance to the basal ganglia. Diverse neuron classes make up striatal microcircuit activity, consisting in the sequential activation of neuronal ensembles. How different neuron classes participate in generating ensemble sequences is unknown. In control mus musculus brain slices in vitro, providing excitatory drive generates ensemble sequences. In Parkinsonian microcircuits captured by a highly recurrent ensemble, a cortical stimulus causes a transitory reconfiguration of neuronal groups alleviating Parkinsonism. Alternation between neuronal ensembles needs interconnectivity, in part due to interneurons, preferentially innervated by incoming afferents. One main class of interneuron expresses parvalbumin (PV+ neurons) and mediates feed-forward inhibition. However, its more global actions within the microcircuit are unknown. Using calcium imaging in ex vivo brain slices simultaneously recording dozens of neurons, we aimed to observe the actions of PV+ neurons within the striatal microcircuit. PV+ neurons in active microcircuits are 5%-11% of the active neurons even if, anatomically, they are

There is a lack of consensus in the international scientific community with respect to the most suitable hydration strategies when attending nulliparous women during low-risk births. This paper describes the protocol for a randomized controlled trial to compare two hydration strategies and their influence on maternal and neonatal morbidity. The study population consists of nulliparous women admitted to the obstetrics department of a University Hospital. The women are being randomized into two groups: the "optimal hydration" group, which will be guaranteed 300 ml/hr liquids (crystalloids and bottled mineral water) with a minimum diuresis of 35 ml/hr; and the "variability in hydration" group, which will receive intravenous (alternating normal saline, Ringer's lactate solution, glucose, or Voluven®) and clear (bottled mineral water or isotonic drinks [Aquarius®]) liquids, without any established perfusion rate, and without established minimum diuresis. Outcomes for mothers include duration of labor, cesarean section, fever, and dehydration. Outcomes for newborns are respiratory distress, hypoglycemia, hyponatremia, jaundice, weight loss over 48 hr, and breastfeeding difficulties. Analysis will be per-protocol. Administering optimal hydration may improve health and safety for mothers and their newborn and reduce maternal and neonatal morbidity. The study is registered at www.clinicaltrials.gov. The project received funding by the Ministry of Health of Spain and is approved by the Research Ethics Committee.

Interactions between plants and their root‐associated fungi (RAF) may influence the relative abundance of tree species and determine forest community diversity. Such plant–soil feedbacks in turn depend on the degree to which spatial distance and phylogenetic relatedness of host trees structure pathogen and mutualist communities, but research detailing these aspects of RAF communities is lacking. Here, we characterize plant–RAF associations across a diverse plant community, focusing on the degree to which RAF communities are structured by spatial distance, host phylogenetic relatedness, and host abundance. We compare results for different functional groups, including both putative mutualists and pathogens, an aspect poorly examined hitherto. We collected roots at regular intervals along ten 50 m by 2 m transects, then used DNA barcoding to identify host plants, and characterize the associated fungal community. Variance partitioning was used to measure the relative contributions of host phylogenetic relatedness and spatial distance to explaining RAF community composition. A weighted linear regression was used to measure the correlation between host abundance and RAF diversity. Phylogenetic distance among hosts was a better predictor of RAF community composition than spatial distance, but this relationship was stronger for putative pathogens than for mutualists, suggesting that pathogens show stronger host preference than mutualists. Across all functional groups, RAF showed similar levels of spatial structure. Additionally, RAF communities of locally abundant plants were less diverse than RAF communities of rare plants. Synthesis. We found that RAF communities are structured by the phylogenetic relatedness of hosts and, to a lesser extent, by spatial distance, with pathogens showing stronger host preference than mutualists. Abundant hosts had less diverse RAF communities than rare hosts, which is notable because abundant plants tend to experience weaker negative plant–soil feedback. Going forward, mechanisms underlying the host abundance‐RAF diversity relationship warrant further investigation. Additionally, the survey approach presented here could be paired with experiments linking RAF community composition to plant recruitment.

Ministerio de Ciencia e Innovacion, Grant/Award Number: FJCI-2017-34967; Sixth Framework Programme, Grant/Award Number: 016181 FOOD, This work was done as part of the IDEFICS study (www.idefics.eu). We gratefully acknowledge the financial support of the European Community within the Sixth RTD Framework Programme Contract No. 016181 (FOOD)., Background Elevated cardiometabolic risk (CMR) is an important factor for cardiovascular diseases later in life while physical fitness seems to decrease CMR. Objective Thus, the aim of the present study is to assess the association between muscular fitness (MF) and cardiorespiratory fitness (CRF) on CMR in European children, both cross-sectional and longitudinally. Methods A total of 289 children (49.5% males) from eight European countries, aged 6 to 9, with longitudinal information on blood pressure, triglycerides, total cholesterol, HDL-cholesterol, homoeostasis model assessment, body mass index, data on fitness level, objectively measured physical activity (PA), diet quality, and total screen time were included. A CMR score was calculated and dichotomized. MF and CRF were also dichotomized. Cross-sectional and longitudinal multilevel logistic regressions adjusting for lifestyle behaviours were performed. Results Reaching a high level of MF during childhood as well as remaining in that level over-time were associated with an 82% and 62% lower probability of high CMR at follow-up, respectively. Also, children who became top CRF over time, showed a 77% lower probability (P < 0.05) of being in the highest CMR quartile at follow-up, independently of sociodemographic and lifestyle indicators. Conclusions A high MF at early childhood and during childhood reduces the odds of having CMR. Same occurs with the improvement of CRF during childhood. These findings highlight the importance of enhancing fitness to avoid CMR already in children., Instituto de Salud Carlos III Spanish Government European Commission FJCI-2017-34967, European Commission 016181 FOOD

Objective This study aimed to test the validity of an automatic oscillometric device to measure the blood pressure (BP) in children (n = 191) and adolescents (n = 127) aged 3 to 18 years. Methods Systolic BP (SBP) and diastolic BP (DBP) levels were measured simultaneous by automatic device and mercury column with Y-connection. To verify the validity, Bland-Altman plots and limits of agreement of 95% (95% LOA), specificity and sensitivity of the device, and the grade of British Hypertension Society (BHS) criteria were used. Results The monitor measurements demonstrated lower measurement bias (mean difference [95% LOA]): 1.4 (-9.9 to 12.8) mmHg in children and 4.3 (-7.8 to 16.5) mmHg in adolescents for SBP. For DBP, it was 2.2 (-7.4 to 11.7) mmHg in children and 1.4 (-8.4 to 11.1) mmHg in adolescents. The sensitivity in children was 21.4 (95% CI = 16.3-26.6), and in adolescents, it was 20.0 (95% CI = 13.2-26.8); the specificity was 95.9 (95% CI = 93.4-98.4) in children and 100.0 (95% CI = 100.0-100.0) in adolescents. The monitor-tested ratings are Grade B for SBP in children and SBP and DBP in adolescents and Grade C for DBP in children. Conclusions The automatic monitor presented high values of specificity and lower values of sensitivity to the diagnosis of HBP; however, it can be considered accurate (lower measurement bias) and valid for epidemiological and clinical practice in accordance with BHS criteria.

Previously, we have shown that chemical excitatory drives such as N-methyl-d-aspartate (NMDA) are capable of activating the striatal microcircuit exhibiting neuronal ensembles that alternate their activity producing temporal sequences. One aim of this work was to demonstrate whether similar activity could be evoked by delivering cortical stimulation. Dynamic calcium imaging allowed us to follow the activity of dozens of neurons with single-cell resolution in mus musculus brain slices. A train of electrical stimuli in the cortex evoked network activity similar to the one induced by bath application of NMDA. Previously, we have also shown that the dopamine-depleted striatal microcircuit increases its spontaneous activity generating dominant recurrent ensembles that interrupt the temporal sequences found in control microcircuits. This activity correlates with parkinsonian pathological activity. Several cortical stimulation protocols such as transcranial magnetic stimulation reduce motor signs of Parkinsonism. Here, we show that cortical stimulation in vitro temporarily eliminates the pathological activity from the dopamine-depleted striatal microcircuit by turning off some neurons that sustain this activity and recruiting new ones that allow transitions between network states, similar to the control circuit. When cortical stimulation is given in the presence of L-DOPA, parkinsonian activity is eliminated during the whole recording period. The present experimental evidence suggests that cortical stimulation such as that generated by transcranial magnetic stimulation, or otherwise, may allow reduce L-DOPA dosage.

The mutualistic interaction between frugivore birds and the fruiting plants they disperse presents an asymmetric interaction pattern, with some species having a more important role (i.e. being essential) for maintaining the structure and functioning of the interaction network. The identification of the biological characteristics of these species is of major importance for the understanding and conservation of seed-dispersal interactions. In this study, I use a network approach and avian seed-dispersal networks from 23 different geographical areas to test five hypotheses about species characteristics determining the structure of the assemblage. I expected (i) large birds to forage on a large number of fruits and (ii) large fruits to be dispersed by few bird species (because of morphological constraints), and (iii) highly energetic fruits to be dispersed by more bird species (in accordance with optimal foraging theory). Besides the number of interacting partners, I also expected (iv) large birds and (v) small energetic fruits to be important for the maintenance of the structure of the interactions in seed-dispersal networks. As species that are closely related are more likely to be similar to each other, I performed phylogenetically corrected analyses to account for this data dependence. Although bird size was not associated to species important in the maintenance of the structure of the seed-dispersal community, I identified that bird species whose diet was strongly dependent on fruits were important for the structure of the network. Regarding the plants, I found that large fruits were dispersed by fewer species, but the most important attribute to predict the role of a fruit was its energy content (higher energy, more bird species dispersing the plant, but low-energy fruits being of conservation concern because they are dispersed by specific species). The results of this study suggest that the role of the species in seed-dispersal assemblages seems to be determined by the role of the species as consumers (frugivory degree for animals) or by their nutritional inputs (energy content for fruits) rather than by morphological constrains.

Species phenotypic traits affect the interaction patterns and the organization of seed‐dispersal interaction networks. Understanding the relationship between species characteristics and network structure help us understand the assembly of natural communities and how communities function. Here, we examine how species traits may affect the rules leading to patterns of interaction among plants and fruit‐eating vertebrates. We study a species‐rich seed‐dispersal system using a model selection approach to examine whether the rules underlying network structure are driven by constraints in fruit resource exploitation, by preferential consumption of fruits by the frugivores, or by a combination of both. We performed analyses for the whole system and for bird and mammal assemblages separately, and identified the animal and plant characteristics shaping interaction rules. The structure of the analyzed interaction network was better explained by constraints in resource exploitation in the case of birds and by preferential consumption of fruits with specific traits for mammals. These contrasting results when looking at bird–plant and mammal–plant interactions suggest that the same type of interaction is organized by different processes depending on the assemblage we focus on. Size‐related restrictions of the interacting species (both for mammals and birds) were the most important factors driving the interaction rules. Our results suggest that the structure of seed‐dispersal interaction networks can be explained using species traits and interaction rules related to simple ecological mechanisms.

Natural wetland ecosystems continue to suffer widespread destruction and degradation. Many recent studies argue that artificial or restored wetlands compensate for wetland loss and are valuable for waterbird conservation. However, detailed comparisons of the value of natural, artificial and restored wetlands are lacking. Our aim was to assess if the restoration or creation of wetlands can fully compensate for the loss of natural wetlands for waterbirds. We compared the waterbird communities in a set of 20 natural, restored and artificial wetlands, all of which are considered important for waterbirds and are located within the same protected area (Donana Natural Space, south-west Spain). We used different measures of diversity, including phylogenetic relatedness, and the proportion of threatened species at national, European and international levels. We found that artificial wetlands have consistently lower value than restored and natural wetlands, with little difference between the latter two. Natural wetlands are essential for conserving diversity across the tree of life and for threatened species, but restored wetlands can be of similar value and can assure maintenance of key ecological processes. Thus, when economic costs per unit area are similar, resources for wetland conservation are better invested in restoration projects than in wetland creation, and caution is required when suggesting that artificial wetlands compensate for the loss of natural wetlands

Aim We aim to characterize the macroecological patterns in the structure of mutualistic seed-dispersal networks. Tropical areas hold more species than temperate ones. This difference in species number may favour ecological processes that minimize interspecific competition in species-rich areas. There is theoretical evidence that both modularity (i.e. the presence of semi-independent groups of highly interacting species) and nestedness (i.e. specialists interact with a subset of the species interacting with generalists) reduce the effects of competition. Thus, we expect high degrees of modularity or nestedness at low latitudes in seed-dispersal networks. Moreover, we test whether climate, topography and human impact influence network structure. Location Thirty-four qualitative and 21 weighted seed-dispersal interaction networks located world-wide. Methods We related the degree of modularity and nestedness of seed-dispersal interaction networks with latitude. To disentangle the macroecological drivers of network structure, we also associated modularity/nestedness with species richness, altitudinal range, human impact and an array of climate predictors: precipitation, temperature, precipitation/temperature seasonality and historical climate-change velocity and anomaly. Results Binary networks showed stronger macroecological patterns than weighted networks. Latitude was unrelated to the structure of seed-dispersal networks, but more nested assemblages were species rich and were located in areas with a high degree of human impact, high temperature seasonality, low precipitation, and, especially on the mainland, high stability in precipitation. Modular networks were species rich and found in areas with low human impact. For both nestedness and modularity, the effects of species richness and human impact were especially strong and consistent. Main conclusions As for previous macroecological studies of mutualistic networks, we found that the structure of seed-dispersal assemblages was related to current and historical climate. The largest influences on nestedness and modularity, however, were the number of competing species and the degree of human impact. This suggests that human disturbance, not just climate, is an important factor determining the structure of a seed-dispersal network.

Split-root system (SRS) approaches allow the differential treatment of separate and independent root systems, while sharing a common aerial part. As such, SRS is a useful tool for the discrimination of systemic (shoot origin) versus local (root/nodule origin) regulation mechanisms. This type of approach is particularly useful when studying the complex regulatory mechanisms governing the symbiosis established between legumes and Rhizobium bacteria. The current work provides an overview of the main insights gained from the application of SRS approaches to understand how nodule number (nodulation autoregulation) and nitrogen fixation are controlled both under non-stressful conditions and in response to a variety of stresses. Nodule number appears to be mainly controlled at the systemic level through a signal which is produced by nodule/root tissue, translocated to the shoot, and transmitted back to the root system, involving shoot Leu-rich repeat receptor-like kinases. In contrast, both local and systemic mechanisms have been shown to operate for the regulation of nitrogenase activity in nodules. Under drought and heavy metal stress, the regulation is mostly local, whereas the application of exogenous nitrogen seems to exert a regulation of nitrogen fixation both at the local and systemic levels.

The key person for the implementation of kindergarten-based behavioural interventions is the kindergarten teacher. When conducting intervention studies in kindergartens, training sessions are needed to train and motivate kindergarten teachers for programme implementation. This paper presents the systematic development of the teachers' trainings executed in the ToyBox-intervention - a kindergarten-based and family-involved obesity prevention programme for children aged 4-6. Based on concepts for the education of kindergarten teachers, on general strategies for successful programme implementation and on the ToyBox programme-specific requirements, the aims of the teachers' trainings were defined and an overall concept was deduced. Regarding the concept for the ToyBox teachers' training sessions, it is concluded that the training modules should focus on presenting information on the practical implementation of the intervention. Furthermore, these modules should also include self-efficacy enhancing components and should give kindergarten teachers opportunities to share experiences. Regarding the didactic methods applied in the ToyBox teachers' training sessions, constructivist learning approaches that facilitate active participation, reflective thinking and personal involvement were implemented. Emphasis was put not only on the content but especially on the didactic methods of teachers' trainings in order to enhance devotion to, and quality and sustainability of the ToyBox-intervention.

Overweight and obesity in children are recognized as a major health problem. The ToyBox-intervention was developed with the aim of preventing obesity in pre-schoolers. Because it is increasingly important to inform policy makers not only on the effects of prevention interventions, but also on their costs and cost-effectiveness, our purpose was to establish a method to estimate the cost-effectiveness of the ToyBox-intervention. In order to estimate the long-term impact of the ToyBox-intervention on health and societal costs, extrapolations of the intervention effect will be conducted to predict children's weight status (based on the body mass index) at adult age. Effects of the adult weight status on the prevalence of obesity-related complications will be modelled through a Markov model, with a total time horizon of 70 years and a cycle length of 1 year. The model will be conducted in six European countries participating in the ToyBox-intervention, based on country-specific economic and epidemiological data. This study describes the methodological rationale and implementation of an analytic model to examine the cost-effectiveness of the ToyBox-intervention for six European countries, in order to inform decision-makers on the value for money of this intervention in the prevention of obesity in pre-schoolers.

Reliable assessments of health-related behaviours are necessary for accurate evaluation on the efficiency of public health interventions. The aim of the current study was to examine the reliability of a self-administered primary caregivers questionnaire (PCQ) used in the ToyBox-intervention. The questionnaire consisted of six sections addressing sociodemographic and perinatal factors, water and beverages consumption, physical activity, snacking and sedentary behaviours. Parents/caregivers from six countries (Belgium, Bulgaria, Germany, Greece, Poland and Spain) were asked to complete the questionnaire twice within a 2-week interval. A total of 93 questionnaires were collected. Test-retest reliability was assessed using intra-class correlation coefficient (ICC). Reliability of the six questionnaire sections was assessed. A stronger agreement was observed in the questions addressing sociodemographic and perinatal factors as opposed to questions addressing behaviours. Findings showed that 92% of the ToyBox PCQ had a moderate-to-excellent test-retest reliability (defined as ICC values from 0.41 to 1) and less than 8% poor test-retest reliability (ICC < 0.40). Out of the total ICC values, 67% showed good-to-excellent reliability (ICC from 0.61 to 1). We conclude that the PCQ is a reliable tool to assess sociodemographic characteristics, perinatal factors and lifestyle behaviours of pre-school children and their families participating in the ToyBox-intervention.

The development of the ToyBox-intervention was based on the outcomes of the preliminary phase of the ToyBox-study, aiming to identify young children's key behaviours and their determinants related to early childhood obesity. The ToyBox-intervention is a multi-component, kindergarten-based, family-involved intervention with a cluster-randomized design, focusing on the promotion of water consumption, healthy snacking, physical activity and the reduction/ breaking up of sedentary time in preschool children and their families. The intervention was implemented during the academic year 2012-2013 in six European countries: Belgium, Bulgaria, Germany, Greece, Poland and Spain. Standardized protocols, methods, tools and material were used in all countries for the implementation of the intervention, as well as for the process, impact, outcome evaluation and the assessment of its cost-effectiveness. A total sample of 7,056 preschool children and their parents/caregivers, stratified by socioeconomic level, provided data during baseline measurements and participated in the intervention. The results of the ToyBox-study are expected to provide a better insight on behaviours associated with early childhood obesity and their determinants and identify effective strategies for its prevention. The aim of the current paper is to describe the design of the ToyBox-intervention and present the characteristics of the study sample as assessed at baseline, prior to the implementation of the intervention.

Process evaluation (PE) is used for the in-depth evaluation of the implementation process of health promotion programmes. The aim of the current paper was to present the PE design and tools used in the ToyBox-intervention. The PE design was based on a three-step approach, including the identification of ToyBox-specific PE elements (step 1), the development of PE tools and harmonization of procedures (step 2), and the implementation of PE using standardized protocol and tools across the intervention countries (step 3). Specifically, to evaluate the implementation of the intervention, teachers' monthly logbooks were recorded (dose delivered, fidelity, dose received); post-intervention questionnaires were completed by parents/caregivers and teachers (dose received); participation and attrition rates were recorded (recruitment, reach); and audit questionnaires and retrospective information on weather conditions were collected (physical and social environment within which the intervention was implemented). Regarding the teachers' training sessions, the researchers who performed the trainings completed evaluation forms and documented teachers' attendance after each training (dose delivered, fidelity, dose received) and teachers completed evaluation forms after each training (dose received). The PE performed in the ToyBox-intervention may contribute in the evaluation of its effectiveness, guide the revision of the intervention material and provide insights for future health promotion programmes and public health policy.

The ToyBox-study aims to develop and test an innovative and evidence-based obesity prevention programme for preschoolers in six European countries: Belgium, Bulgaria, Germany, Greece, Poland and Sp ...

Symbiotic nitrogen fixation is one of the first physiological processes inhibited in legume plants under water-deficit conditions. Despite the progress made in the last decades, the molecular mechanisms behind this regulation are not fully understood yet. Recent proteomic work carried out in the model legume Medicago truncatula provided the first indications of a possible involvement of nodule methionine (Met) biosynthesis and related pathways in response to water-deficit conditions. To better understand this involvement, the drought-induced changes in expression and content of enzymes involved in the biosynthesis of Met, S-adenosyl-L-methionine (SAM) and ethylene in M.truncatula root and nodules were analyzed using targeted approaches. Nitrogen-fixing plants were subjected to a progressive water deficit and a subsequent recovery period. Besides the physiological characterization of the plants, the content of total sulphur, sulphate and main S-containing metabolites was measured. Results presented here show that S availability is not a limiting factor in the drought-induced decline of nitrogen fixation rates in M.truncatula plants and provide evidences for a down-regulation of the Met and ethylene biosynthesis pathways in roots and nodules in response to water-deficit conditions.

Wetland restoration is increasingly important to reverse habitat degradation, recover ecosystem services, and maintain biodiversity. To aid project design, more information is required on the influence of wetland size, depth of water, and isolation on the waterbird communities that become established. During a restoration project in Donana, one of the Europe's most important wetland complexes, an experimental network of 96 temporary ponds with standard shape but variable size, water depth, and isolation was created. We surveyed the waterbird community in spring from 2006 to 2008 and related species abundance and richness to abiotic pond characteristics. We also performed analyses pooling species by foraging guilds or body size. Waterbird abundance and species richness were highest in 2007, the wettest year when the ponds had longer hydroperiods. Larger ponds had consistently higher abundance and species richness for the entire community and for different guilds and body sizes. Moreover, the density of birds per hectare was higher in large ponds than in the smaller ones. Pond isolation and excavated depth did not affect overall abundance and richness, although opposing effects of depth were observed on some size classes, and ducks and large birds preferred isolated ponds. Some bird groups preferred ponds at a particular location. This is the first waterbird study to address the importance of pond size, depth, and isolation independently of confounding variables such as pond shape. It illustrates the varied responses from different bird groups and demonstrates the importance of varying depth, location, and isolation to enhance community abundance and diversity.

Summary 1. A long-standing question in ecology is how natural populations respond to a changing environment. Emergent optimal foraging theory-based models for individual variation go beyond the population level and predict how its individuals would respond to disturbances that produce changes in resource availability. 2. Evaluating variations in resource use patterns at the intrapopulation level in wild populations under changing environmental conditions would allow to further advance in the research on foraging ecology and evolution by gaining a better idea of the underlying mechanisms explaining trophic diversity. 3. In this study, we use a large spatio-temporal scale data set (western continental Europe, 1968–2006) on the diet of Bonelli’s Eagle Aquila fasciata breeding pairs to analyse the predator trophic responses at the intrapopulation level to a prey population crash. In particular, we borrow metrics from studies on network structure and intrapopulation variation to understand how an emerging infectious disease [the rabbit haemorrhagic disease (RHD)] that caused the density of the eagle’s primary prey (rabbit Oryctolagus cuniculus) to dramatically drop across Europe impacted on resource use patterns of this endangered raptor. 4. Following the major RHD outbreak, substantial changes in Bonelli’s Eagle’s diet diversity and organisation patterns at the intrapopulation level took place. Dietary variation among breeding pairs was larger after than before the outbreak. Before RHD, there were no clusters of pairs with similar diets, but significant clustering emerged after RHD. Moreover, diets at the pair level presented a nested pattern before RHD, but not after. 5. Here, we reveal how intrapopulation patterns of resource use can quantitatively and qualitatively vary, given drastic changes in resource availability. 6. For the first time, we show that a pathogen of a prey species can indirectly impact the intrapopulation patterns of resource use of an endangered predator.

The ideal free distribution (IFD) model predicts that a density-dependent mechanism operates to regulate habitat selection and reproductive performance. We studied a Little Grebe Tachybaptus ruficollis population, which breeds on irrigation ponds in the Vega Baja Valley (southeastern Spain) to test the premises of the IFD model. These ponds are highly dynamic because they are managed according to agricultural requirements, and are subject to different levels of disturbance, which can change the quality of individual ponds across the landscape. Surveys were carried out during the breeding season from 2002 to 2006, with reproduction performance estimated during two consecutive breeding seasons, 2003 and 2004. Occupation frequency differed from random, indicating preference for some ponds over others. Habitat features such as pond construction and design, the presence of submerged vegetation, vegetation along the shore and reed beds, and pond area correlated with occupation frequency and might be considered to be indicators of pond quality. Ponds were occupied sequentially from best to worst. Thus, when the population size increased, the number of low-quality ponds occupied also increased. High-quality ponds held more breeding pairs than low-quality ones, resulting in the mean reproductive success per breeding pair being independent of pond quality. Little Grebes therefore occupy ponds in a manner consistent with the expectations of the IFD model.

The incidence of intentional or accidental valproic acid (VPA) overdose is increasing. Severe VPA toxicity may lead to coma and death. Traditionally the treatment of patients with VPA toxicity has been limited to supportive measures. VPA is highly protein bound and therefore it is considered not to be removable by extracorporeal means. However, studies of VPA toxicokinetics indicate that at blood levels that exceed therapeutic concentrations, VPA protein binding sites become saturated, leading to increased concentration of the free unbound drug. The free unbound drug has a small molecular weight and therefore it is theoretically amenable to removal by extracorporeal means. We present a patient with VPA toxicity who was successfully treated with "in-series" hemodialysis and hemoperfusion followed by continuous venovenous hemodiafiltration (CVVHDF) and review the literature on the management of VPA toxicity using extracorporeal therapies.

Hyperparathyroidism and alterations in bone and mineral metabolism are known to occur early in the course of chronic kidney disease (CKD). The etiology of these abnormalities is multifactorial. Phosphate retention and alterations in vitamin D metabolism play important roles. The recent practice guidelines from the National Kidney Foundation (NKF) have emphasized the need to approach these problems early in the course of CKD by measuring the levels of parathyroid hormone when glomerular filtration rates (GFRs) fall to less than 60 ml/min. If hyperparathyroidism is detected, then treatment must be undertaken to try to prevent its progression. Strict guidelines have been proposed for the management of these alterations in bone and mineral metabolism when kidney disease progresses to CKD stage V. Although the initial recommendations were often opinion based, evidence is accumulating to support these views. However, with our current therapeutic armamentarium, there is considerable difficulty in achieving these practice guidelines for calcium, phosphorus, calcium-phosphorus product, and parathyroid hormone on a sustained basis. New therapeutic agents are becoming available that will help with phosphorus control and control of hyperparathyroidism, and minimize the calcium load. Additional beneficial effects of vitamin D analogs are being uncovered and are being intensively investigated. It is hoped that with attention to these practice guidelines, control of the abnormalities of mineral metabolism will be improved and will lead to improved patient outcomes.

Derangements in mineral metabolism are known to occur early in the course of chronic kidney disease (CKD). Recent clinical practice guidelines are designed to focus on the problem early in the course of kidney disease, when it is recommended to evaluate the levels of parathyroid hormone (PTH) and to try to intervene early if the levels are elevated. To begin early intervention for hyperparathyroidism in chronic kidney disease will require involvement of primary care physicians and other subspecialty groups to identify the patients at risk and begin to intervene with measures to control hyperparathyroidism and its consequences on mineral metabolism. It has recently been demonstrated that chronic kidney disease is a significant risk factor for vitamin D deficiency and since abnormalities in vitamin D metabolism are important in the generation of hyperparathyroidism, this is an issue that needs direct attention. Studies are needed to assess the effects of correcting this vitamin D deficiency in early CKD. As kidney disease progresses, efforts to control hyperparathyroidism will likely need to be intensified and several therapeutic options are available, such as phosphate binders, repletion of vitamin D, the use of active vitamin D sterols, or the use of vitamin D analogs. In addition, it is important to define the appropriate PTH values that need to be achieved to minimize complications on bone. Such studies are in progress at the present time to validate the current more specific PTH assays. Strict guidelines have been proposed for the management of bone and mineral metabolism in patients with CKD stage V on dialysis, and although these challenging recommendations were initially opinion-based, there is mounting evidence which provides confirmation of these targets as relevant. Treatment options for patients on dialysis involve the full spectrum of agents which include phosphate binders, active vitamin D sterols (often given parenterally), the use of calcimimetic agents, surgical parathyroidectomy, and evaluation of appropriate levels of dialysate calcium. Similar to early stages of CKD, studies are in progress to refine the PTH targets with the newer PTH assays. With increased focus on the complications of bone and mineral metabolism as part of the continuum of chronic kidney disease, and with a variety of new therapies available, it is anticipated that improved patient outcomes should be achievable in this patient group.

Semi-leafless varieties of pea have considerable agronomic importance and it has been suggested that they may have a superior response to water deficits than conventional varieties. However, these varieties are poorly characterized from a physiological point of view and there is lack of a physiological basis for their supposed better performance under conditions of water deficit. Here, we describe the solute distribution in the different leaf structures of a semi-leafless pea variety (Pisum sativum L.) under non-limiting water conditions and under water stress. A conventional variety was subjected to the same conditions for comparative purposes. A detailed study was carried out both at the tissue level and at the single cell level. In control conditions, epidermal vacuoles of tendrils showed a different ion distribution of those of the laminar leaf structures. However, under water deficit, only stipules of the semi-leafless variety showed a significantly higher capability to increase osmolarity. This occurred by accumulating potassium, magnesium and chloride to a higher extent than other leaf structures. The inability of performing an adequate osmotic adjustment in tendrils may be the cause of the lack of a better response to water deficit.

Recent research has shown that nodule nitrogen fixation is limited under a wide range of environmental constraints by lowered carbon flux within the nodule due to down-regulation of sucrose synthase activity. The aim of this work was to elucidate whether an increase in both carbon flux and activity of enzymes of carbon metabolism in nodules may lead to an increased nitrogen fixation. We report the effects caused by a continuous exposure to atmospheric CO 2 enrichment in modulated pea plants. CO 2 enrichment led to an enhanced whole-plant growth and increased nodule biomass. Moreover, nodules of plants grown at increased CO 2 showed a higher sugar content as well as enhancement of some activities related to nodule carbon metabolism, such as sucrose synthase, UDP glucose pyrophosphorylase and phosphoenolpyruvate carboxylase. Indeed, acetylene reduction activity, measured by the classical technique, was increased more than four times. However, when specific nitrogen fixation was determined as hydrogen evolution, no significant differences were detected, consistent with the lack of changes of enzymes involved in nitrogen metabolism such as glutamate synthase and aspartate aminotransferase. These results are discussed in the context of the regulation of nitrogen fixation and nodule metabolism.

The recessive gene af produces a modification of Pisum sativum L. leaf morphology, where leaflets are replaced by tendrils. Previous reports have suggested that tendrils may contribute to plant growth in a similar way to flat leaf structures, but these reports have been restricted to carbon metabolism. In this work, we investigate the nitrate reduction (nitrate reductase activity) capacity of tendrils of a semi-leafless variety, Solara, in relation to other leaf structures. Maximum nitrate reductase activity (EC 1.6.6.1), expressed on a protein basis, was significantly lower in tendrils compared to flat structures. However, the activation state of nitrate reductase was significantly higher in tendrils, reaching 70%, compared to flat leaf structures. According to these results, tendrils contributed up to 25% of the overall plant nitrate reduction in the semi-leafless variety. This figure was even higher when nitrate reduction was calculated from in vivo measurements. The results are discussed in relation to nitrate, magnesium, carbohydrates, amino acids and adenylate levels of tendrils.

1. The breakdown of oak (Quercus robur L.), chestnut (Castanea sativa Miller) and eucalypt (Eucalyptus globulus Labill.) litter enclosed in 5-mm mesh bags was compared between first-order headwaters (two with native riparian forest and two with eucalypt plantations) and a third-order reach of Aguera stream. Weight loss and dynamics of phosphorus and nitrogen in litter were studied for a period of 155 days. 2. Among the different sites, processing rates ranged from 0.0045 to 0.0080 day–1 for chestnut leaf litter, from 0.0036 to 0.0051 day–1 for oak, and from 0.0027 to 0.0158 day–1 for eucalypt. 3. The availability of nutrients in water clearly influenced nitrogen and phosphorus dynamics in litter. In headwater reaches, net immobilization was not observed and losses of phosphorus and nitrogen followed mass loss. However, there was an enrichment of litter at the low reach, where influence of human settlements—located upstream—could lead to a greater availability of phosphorus in water. 4. The enhancement of litter decay by the exogenous nutrient supply depended on leaf quality, as only the processing rate of eucalypt increased at the nutrient-rich site. 5. The processing rates differed little among headwaters, suggesting that riparian forest type, i.e. deciduous forest v eucalypt plantations, did not affect litter decay in the stream.

Valproic acid toxicity is not infrequent and is difficult to treat, as there is no specific antidote. In the past, this drug was considered unremovable by extracorporeal methods because of the highly protein bound state (90–95%). However, recent reports suggest that the toxicokinetics of valproate vary considerably from the pharmacokinetics at therapeutic levels, and at higher concentrations protein-binding sites become saturated. The drug's relatively low molecular weight (144 Da), small volume of distribution (0.13–0.23 L/kg), and saturable protein binding render it potentially amenable to extracorporeal removal, but published experience is scant and there are only a few reported instances in which patients were successfully treated with extracorporeal methods. Here we report the case of a patient with serious valproate toxicity treated with simultaneous “in series” hemodialysis and hemoperfusion followed by continuous veno-venous hemodiafiltration. A 35-year-old homeless male presented to the emergency department after ingesting 120 pills of valproic acid. Initial valproic acid level was 59 µg/mL. Urine drug screen was negative and serum chemistries including LFTs were normal. He was treated with activated charcoal and admitted to the intensive care unit. 4 h later, he developed respiratory failure and became hemodynamically unstable. He was intubated and successfully resuscitated. Subsequently, the valproic acid level was found to be 553 µg/mL and a decision was made to employ extracorporeal methods for drug removal. “In series” hemodialysis and hemoperfusion were done for 4 h and simultaneous blood samples for measurement of valproic acid levels were obtained as blood entered the hemoperfusion column (arterial) and as it exited the hemodialysis membrane (venous). Extraction ratio, whole blood, and plasma clearances were calculated and they compared substantially with the published data. The above measures decreased valproate levels from 572.6 (pre-dialysis level) to 203.2 µg/mL. This was followed by continuous veno-venous hemodiafiltration for 18 h, which sustained the drug removal and prevented significant post-dialytic rebound. This is the first reported instance in which these three methods were administered together successfully in the management of valproic acid toxicity.