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The application of anaerobic digestion technology is growing worldwide because of its economic and environmental benefits. As a consequence, a number of studies and research activities dealing with the determination of the biogas potential of solid organic substrates have been carrying out in the recent years. Therefore, it is of particular importance to define a protocol for the determination of the ultimate methane potential for a given solid substrates. In fact, this parameter determines, to a certain extent, both design and economic details of a biogas plant. Furthermore, the definition of common units to be used in anaerobic assays is increasingly requested from the scientific and engineering community. This paper presents some guidelines for biomethane potential assays prepared by the Task Group for the Anaerobic Biodegradation, Activity and Inhibition Assays of the Anaerobic Digestion Specialist Group of the International Water Association. This is the first step for the definition of a standard protocol.

Summary Background Previous studies have demonstrated significant variability in the processes of care and outcomes of chronic obstructive pulmonary disease (COPD) exacerbations. The AUDIPOC is a Spanish nationwide clinical audit that identified large between‐hospital variations in care and clinical outcomes. Here, we test the hypothesis that these variations can be attributed to either patient characteristics, hospital characteristics and/or the so‐called hospital‐clustering effect, which indicates that patients with similar characteristics may experience different processes of care and outcomes depending on the hospital to which they are admitted. Methods A clinical audit of 5178 COPD patients consecutively admitted to 129 Spanish public hospitals was performed, with a 90‐day follow‐up. Multilevel regression analysis was conducted to model the probability of patients experiencing adverse outcomes. For each outcome, an empty model (with no independent variables) was fitted to assess the clustering effect, followed by a model adjusted for the patient‐ and hospital‐level covariables. The hospital‐clustering effect was estimated using the intracluster correlation coefficient (ICC); the cluster heterogeneity was estimated with the median odds ratio (MOR), and the coefficients of predictors were estimated with the odds ratio (OR). Results In the empty models, the ICC (MOR) for inpatient mortality and the follow‐up mortality and readmission were 0.10 (1.80), 0.08 (1.65) and 0.01 (1.24), respectively. In the adjusted models, the variables that most represented the patients’ clinical conditions and interventions were identified as outcome predictors and further reduced the hospital variations. By contrast, the resource factors were primarily unrelated with outcomes. Conclusions This study demonstrates a noteworthy reduction in the observed crude between‐hospital variation in outcomes after accounting for the hospital‐cluster effect and the variables representing patient's clinical conditions. This emphasises the predictor importance of the patients’ clinical conditions and interventions, and understates the impacts of hospital resources and organisational factors.

Dengue and Zika are two of the most important human viral pathogens worldwide. In both cases, the envelope glycoprotein E is the main target of the antibody response. Recently, new complex quaternary epitopes were identified which are the consequence of the arrangement of the antiparallel E dimers on the viral surface. Such epitopes can be exploited to develop more efficient cross-neutralizing vaccines. Here we describe a successful covalent stabilization of E dimers from Dengue and Zika viruses in mammalian cells. Folding and dimerization of secretory E was found to be strongly dependent on temperature but independent of PrM co-expression. In addition, we found that, due to the close relationship between flaviviruses, Dengue and Zika viruses E proteins can form heterodimers and assemble into mosaic viral particles. Finally, we present new virus-free analytical platforms to study and screen antibody responses against Dengue and Zika, which allow for differentiation of epitopes restricted to specific domains, dimers and higher order arrangements of E.

Anxiety disorders affect millions of people worldwide and impair health, happiness, and productivity on a massive scale. Developmental research points to a connection between early-life behavioral inhibition and the eventual development of these disorders. Our group has previously shown that measures of behavioral inhibition in young rhesus monkeys ( Macaca mulatta ) predict anxiety-like behavior later in life. In recent years, clinical and basic researchers have implicated the central extended amygdala (EAc)—a neuroanatomical concept that includes the central nucleus of the amygdala (Ce) and the bed nucleus of the stria terminalis (BST)—as a key neural substrate for the expression of anxious and inhibited behavior. An improved understanding of how early-life behavioral inhibition relates to an increased lifetime risk of anxiety disorders—and how this relationship is mediated by alterations in the EAc—could lead to improved treatments and preventive strategies. In this study, we explored the relationships between infant behavioral inhibition and peri-adolescent defensive behavior and brain metabolism in 18 female rhesus monkeys. We coupled a mildly threatening behavioral assay with concurrent multimodal neuroimaging, and related those findings to various measures of infant temperament. To score the behavioral assay, we developed and validated UC-Freeze , a semi-automated machine-learning (ML) tool that uses unsupervised clustering to quantify freezing. Consistent with previous work, we found that heightened Ce metabolism predicted elevated defensive behavior (i.e., more freezing) in the presence of an unfamiliar human intruder. Although we found no link between infant-inhibited temperament and peri-adolescent EAc metabolism or defensive behavior, we did identify infant nervous temperament as a significant predictor of peri-adolescent defensive behavior. Our findings suggest a connection between infant nervous temperament and the eventual development of anxiety and depressive disorders. Moreover, our approach highlights the potential for ML tools to augment existing behavioral neuroscience methods.

The establishment of a region of suppressed recombination is a critical change during sex chromosome evolution, leading to such properties as Y (and W) chromosome genetic degeneration, accumulation of repetitive sequences and heteromorphism. Although chromosome inversions can cause large regions to have suppressed recombination, and inversions are sometimes involved in sex chromosome evolution, gradual expansion of the non-recombining region could potentially sometimes occur. We here test whether closer linkage has recently evolved between the sex-determining region and several genes that are partially sex-linked in Silene latifolia, using Silene dioica, a closely related dioecious plants whose XY sex chromosome system is inherited from a common ancestor. The S. latifolia pseudoautosomal region (PAR) includes several genes extremely closely linked to the fully Y-linked region. These genes were added to an ancestral PAR of the sex chromosome pair in two distinct events probably involving translocations of autosomal genome regions causing multiple genes to become partially sex-linked. Close linkage with the PAR boundary must have evolved since these additions, because some genes added in both events now show almost complete sex linkage in S. latifolia. We compared diversity patterns of five such S. latifolia PAR boundary genes with their orthologues in S. dioica, including all three regions of the PAR (one gene that was in the ancestral PAR and two from each of the added regions). The results suggest recent recombination suppression in S. latifolia, since its split from S. dioica.

A paradigm in paleoclimatology holds that shifts in the mean position of the Intertropical Convergence Zone were the dominant climatic mechanism controlling rainfall in the tropics during the last glacial period. We present a new paleo-rainfall reconstruction based on speleothem stable oxygen isotopes record from Colombia, which spans most of the last glacial cycle. The strength and positioning of the Intertropical Convergence Zone over northern South America were more strongly affected by summer insolation at high northern latitudes than by local insolation during the last glacial cycle, resulting in an antiphased relationship with climate in the Cariaco Basin. Our data also provide new insight into how orbital forcing amplified/dampened Intertropical Convergence Zone precipitation during millennial-scale events. During Greenland Stadial events, the Intertropical Convergence Zone was positioned close to the latitude of El Peñon, as expressed by more negative δ 18 O values. Greenland Interstadial events are marked by relatively high stable oxygen isotope values and reduced rainfall in the El Peñon record, suggesting a northward withdrawal of the Intertropical Convergence Zone. During some Heinrich Stadial events, and especially Heinrich Stadial 1, the Intertropical Convergence Zone must have been displaced away from its modern location near El Peñon, as conditions were very dry at both El Peñon and Cariaco.

The anammox-based process ELAN® was started-up in two different sequencing batch reactor (SBR) pilot plant reactors treating municipal anaerobic digester supernatant. The main difference in the operation of both reactors was the dissolved oxygen (DO) concentration in the bulk liquid. SBR-1 was started at a DO value of 0.4 mg O2/L whereas SBR-2 was started at DO values of 3.0 mg O2/L. Despite both reactors working at a nitrogen removal rate of around 0.6 g N/(L d), in SBR-1, granules represented only a small fraction of the total biomass and reached a diameter of 1.1 mm after 7 months of operation, while in SBR-2 the biomass was mainly composed of granules with an average diameter of 3.2 mm after the same operational period. Oxygen microelectrode profiling revealed that granules from SBR-2 where only fully penetrated by oxygen with DO concentrations of 8 mg O2/L while granules from SBR-1 were already oxygen penetrated at DO concentrations of 1 mg O2/L. In this way granules from SBR-2 performed better due to the thick layer of ammonia oxidizing bacteria, which accounted for up to 20% of all the microbial populations, which protected the anammox bacteria from non-suitable liquid media conditions.

The feasibility of treating the supernatant of a municipal sludge digester supplemented with co-substrates by means of an anammox-based process (ELAN®) was tested in Guillarei (NW of Spain). Ammonia concentration measured in the supernatant of the sludge digester varied in the range 800–1,500 g N/m3 due to the fact that the sludge produced in the plant was co-digested with wastes coming from surrounding food industries. Treating this supernatant in the ELAN® reactor, nitrogen removal rates up to 1.1 kg N/(m3 d) were reached in experiments run in a pilot plant reactor operated in batch mode. No nitrite oxidation was registered after several months of operation despite the average dissolved oxygen (DO) concentrations being 1.5 g O2/m3 and the temperature reaching values as low as 18 °C. By keeping the DO set point at 1–2 g O2/m3 and tuning the hydraulic retention time, the stability of the process was guaranteed and the presence of co-substrates in the anaerobic digester did not affect negatively the operation of the autotrophic nitrogen removal process. Due to the success of the pilot plant experiment, an upscale of the process to full scale is proposed. Mass balances applied to Guillarei wastewater treatment plant revealed that in the main stream line the average denitrification rate calculated with the data of year 2011 was 226 kg N/d. Since the nitrogen removal efficiency is limited by the amount of readily biodegradable organic matter available to carry out denitrification in the water line, the implementation of an anammox-based process to treat the supernatant seems the best option to improve the effluent quality in terms of nitrogen content. The nitrogen removal rate in the sludge line would be 30 times higher than the one in the water line. The implementation of the process would improve the energetic balance and the nitrogen removal performance of the plant.

A fabrication method is developed to produce a microfluidic device to test cell adhesion to polymeric materials. The process is able to produce channels with walls of any spin coatable polymer. The method is a modification of the existing poly-dimethylsiloxane soft lithography method and, therefore, it is compatible with sealing methods and equipment of most microfluidic laboratories. The molds are produced by xurography, simplifying the fabrication in laboratories without sophisticated equipment for photolithography. The fabrication method is tested by determining the effective differences in bacterial adhesion in five different materials. These materials have different surface hydrophobicities and charges. The major drawback of the method is the location of the region of interest in a lowered surface. It is demonstrated by bacterial adhesion experiments that this drawback has a negligible effect on adhesion. The flow in the device was characterized by computational fluid dynamics and it was shown that shear stress in the region of interest can be calculated by numerical methods and by an analytical equation for rectangular channels. The device is therefore validated for adhesion tests.