Chemical characteristics and valuation of sewage sludge from four different wastewater treatment plants.

Sewage sludge contains plant nutrients and organic matter in its composition, making it a potential partial substitute for mineral fertilizers if it meets environmental, agronomic, and sanitary standards. The objective was to evaluate the content of nutrients and heavy metals in the sludge generated in four wastewater treatment stations (WWTPs) in Rio de Janeiro state and assess its potential value and usefulness. The samples of 19 batches from the WTTPs Alegria, Barra da Tijuca, Ilha do Governador, and Sarapuí were analyzed. The WWTPs differ in methods and processes used for treating sewage and sludge. The total contents of C, N, P, K, Ca, Mg, Fe, Al, Na, Co, Mn, As, Ba, Cd, Cr, Cu, Ni, Se, Pb, and Zn were evaluated, as well as the ratio C/N, pH, organic matter content, and electrical conductivity. The grouping of sludge samples was assessed using principal components (PCA) and cluster analysis. The economic valuation of sludge was conducted utilizing the substitute goods method, which compared the sludge's N-P-K contents with the prices of consolidated nutrient sources. All the evaluated sludge batches exhibited concentrations of heavy metals below the limits allowed by Brazilian law, along with high levels of nutrients and organic matter. Considering the chemical characteristics, all evaluated materials showed potential for agricultural use, but it is crucial to evaluate the microbiological characteristics of sludge batches before agriculture application. PCA and cluster analysis demonstrated that sludge samples from the same WWTP clustered close to each other, demonstrating higher similarity among themselves than with samples from other WWTPs. The sludge had an average added value of U$ 88.46 per megagram, considering the total contents of N, P, and K in its composition. Land application of sewage sludge can reduce the need to purchase mineral fertilizers, thereby supporting the feasibility of reusing this material in the agricultural sector.

Structure of a 30S pre-initiation complex stalled by GE81112 reveals structural parallels in bacterial and eukaryotic protein synthesis initiation pathways.

In bacteria, the start site and the reading frame of the messenger RNA are selected by the small ribosomal subunit (30S) when the start codon, typically an AUG, is decoded in the P-site by the initiator tRNA in a process guided and controlled by three initiation factors. This process can be efficiently inhibited by GE81112, a natural tetrapeptide antibiotic that is highly specific toward bacteria. Here GE81112 was used to stabilize the 30S pre-initiation complex and obtain its structure by cryo-electron microscopy. The results obtained reveal the occurrence of changes in both the ribosome conformation and initiator tRNA position that may play a critical role in controlling translational fidelity. Furthermore, the structure highlights similarities with the early steps of initiation in eukaryotes suggesting that shared structural features guide initiation in all kingdoms of life.

RsgA couples the maturation state of the 30S ribosomal decoding center to activation of its GTPase pocket.

During 30S ribosomal subunit biogenesis, assembly factors are believed to prevent accumulation of misfolded intermediate states of low free energy that slowly convert into mature 30S subunits, namely, kinetically trapped particles. Among the assembly factors, the circularly permuted GTPase, RsgA, plays a crucial role in the maturation of the 30S decoding center. Here, directed hydroxyl radical probing and single particle cryo-EM are employed to elucidate RsgA΄s mechanism of action. Our results show that RsgA destabilizes the 30S structure, including late binding r-proteins, providing a structural basis for avoiding kinetically trapped assembly intermediates. Moreover, RsgA exploits its distinct GTPase pocket and specific interactions with the 30S to coordinate GTPase activation with the maturation state of the 30S subunit. This coordination validates the architecture of the decoding center and facilitates the timely release of RsgA to control the progression of 30S biogenesis.

Phosphorylation Alters the Residual Structure and Interactions of the Regulatory L1 Linker Connecting NBD1 to the Membrane-Bound Domain in SUR2B.

ATP-sensitive potassium (KATP) channels in vascular smooth muscle are comprised of four pore-forming Kir6.1 subunits and four copies of the sulfonylurea receptor 2B (SUR2B), which acts as a regulator of channel gating. Recent electron cryo-microscopy (cryo-EM) structures of the pancreatic KATP channel show a central Kir6.2 pore that is surrounded by the SUR1 subunits. Mutations in the L1 linker connecting the first membrane-spanning domain and the first nucleotide binding domain (NBD1) in SUR2B cause cardiac disease; however, this part of the protein is not resolved in the cryo-EM structures. Phosphorylation of the L1 linker, by protein kinase A, disrupts its interactions with NBD1, which increases the MgATP affinity of NBD1 and KATP channel gating. To elucidate the mode by which the L1 linker regulates KATP channels, we have probed the effects of phosphorylation on its structure and interactions using nuclear magnetic resonance (NMR) spectroscopy and other techniques. We demonstrate that the L1 linker is an intrinsically disordered region of SUR2B but possesses residual secondary and compact structure, both of which are disrupted with phosphorylation. NMR binding studies demonstrate that phosphorylation alters the mode by which the L1 linker interacts with NBD1. The data show that L1 linker residues with the greatest α-helical propensity also form the most stable interaction with NBD1, highlighting a hot spot within the L1 linker. This hot spot is the site of disease-causing mutations and is associated with other processes that regulate KATP channel gating. These data provide insights into the mode by which the phospho-regulatory L1 linker regulates KATP channels.

Phosphorylation-dependent changes in nucleotide binding, conformation, and dynamics of the first nucleotide binding domain (NBD1) of the sulfonylurea receptor 2B (SUR2B).

The sulfonylurea receptor 2B (SUR2B) forms the regulatory subunit of ATP-sensitive potassium (KATP) channels in vascular smooth muscle. Phosphorylation of the SUR2B nucleotide binding domains (NBD1 and NBD2) by protein kinase A results in increased channel open probability. Here, we investigate the effects of phosphorylation on the structure and nucleotide binding properties of NBD1. Phosphorylation sites in SUR2B NBD1 are located in an N-terminal tail that is disordered. Nuclear magnetic resonance (NMR) data indicate that phosphorylation of the N-terminal tail affects multiple residues in NBD1, including residues in the NBD2-binding site, and results in altered conformation and dynamics of NBD1. NMR spectra of NBD1 lacking the N-terminal tail, NBD1-ΔN, suggest that phosphorylation disrupts interactions of the N-terminal tail with the core of NBD1, a model supported by dynamic light scattering. Increased nucleotide binding of phosphorylated NBD1 and NBD1-ΔN, compared with non-phosphorylated NBD1, suggests that by disrupting the interaction of the NBD core with the N-terminal tail, phosphorylation also exposes the MgATP-binding site on NBD1. These data provide insights into the molecular basis by which phosphorylation of SUR2B NBD1 activates KATP channels.

Structural characterization of an alternative mode of tigecycline binding to the bacterial ribosome.

Although both tetracycline and tigecycline inhibit protein synthesis by sterically hindering the binding of tRNA to the ribosomal A site, tigecycline shows increased efficacy in both in vitro and in vivo activity assays and escapes the most common resistance mechanisms associated with the tetracycline class of antibiotics. These differences in activities are attributed to the tert-butyl-glycylamido side chain found in tigecycline. Our structural analysis by X-ray crystallography shows that tigecycline binds the bacterial 30S ribosomal subunit with its tail in an extended conformation and makes extensive interactions with the 16S rRNA nucleotide C1054. These interactions restrict the mobility of C1054 and contribute to the antimicrobial activity of tigecycline, including its resistance to the ribosomal protection proteins.

Geographical and climatic factors and depression risk in the SUN project.

BACKGROUND: Depression incidence has been related with seasonal periodicity and climate. The aim of the study was to estimate the possible association between depression and specific meteorological factors, namely temperature, light and rain. METHODS: In total, 13,938 participants from the SUN (Seguimiento Universidad de Navarra) cohort study were included in the analysis. Subjects were classified according to daily mean temperature, number of daylight hours and amount of rain, by year, at their geographical area of residence, data supplied by the Spanish Agency of Meteorology. Participants were considered as incident cases of depression whenever they reported a physician diagnosis of depression or the use of antidepressant medication in any of the follow-up questionnaires. Cox regression models were fit to assess the relationship between climatic and geographical factors and the incidence of depression. RESULTS: Male subjects living in the south and centre areas of Spain showed a higher risk to develop depression compared with those living in the north area (hazard ratio = 1.6, 95% CI = 1.16-2.23 and hazard ratio = 1.41, 95% CI = 1.06-1.87, respectively). Moreover, among males, a direct association between the number of daily light hours and mean temperature and the risk of depression was also found. For men, living in rainy areas was associated with a lower risk of developing depression. CONCLUSION: Our results suggest that climate-depression relationship is more complex than previously thought, and strongly different between men and women.

Updating functional brain units: Insights far beyond Luria.

This paper reviews Luria's model of the three functional units of the brain. To meet this objective, several issues were reviewed: the theory of functional systems and the contributions of phylogenesis and embryogenesis to the brain's functional organization. This review revealed several facts. In the first place, the relationship/integration of basic homeostatic needs with complex forms of behavior. Secondly, the multi-scale hierarchical and distributed organization of the brain and interactions between cells and systems. Thirdly, the phylogenetic role of exaptation, especially in basal ganglia and cerebellum expansion. Finally, the tripartite embryogenetic organization of the brain: rhinic, limbic/paralimbic, and supralimbic zones. Obviously, these principles of brain organization are in contradiction with attempts to establish separate functional brain units. The proposed new model is made up of two large integrated complexes: a primordial-limbic complex (Luria's Unit I) and a telencephalic-cortical complex (Luria's Units II and III). As a result, five functional units were delineated: Unit I. Primordial or preferential (brainstem), for life-support, behavioral modulation, and waking regulation; Unit II. Limbic and paralimbic systems, for emotions and hedonic evaluation (danger and relevance detection and contribution to reward/motivational processing) and the creation of cognitive maps (contextual memory, navigation, and generativity [imagination]); Unit III. Telencephalic-cortical, for sensorimotor and cognitive processing (gnosis, praxis, language, calculation, etc.), semantic and episodic (contextual) memory processing, and multimodal conscious agency; Unit IV. Basal ganglia systems, for behavior selection and reinforcement (reward-oriented behavior); Unit V. Cerebellar systems, for the prediction/anticipation (orthometric supervision) of the outcome of an action. The proposed brain units are nothing more than abstractions within the brain's simultaneous and distributed physiological processes. As function transcends anatomy, the model necessarily involves transition and overlap between structures. Beyond the classic approaches, this review includes information on recent systemic perspectives on functional brain organization. The limitations of this review are discussed.

Effect of zinc intake on mental and motor development in infants: a meta-analysis.

A systematic review and meta-analysis of available randomized controlled trials (RCTs) was conducted to evaluate the effect of zinc (Zn) intake on mental and motor development in infants. Out of 5500 studies identified through electronic searches and reference lists, 5 RCTs were selected after applying the exclusion/inclusion criteria. The influence of Zn intake on mental and motor development was considered in the overall meta-analysis. Other variables were also taken into account as possible effect modifiers: doses of Zn intake, intervention duration, nutritional situation, and risk of bias. Indices of mental and motor development assessed were the Mental Development Index (MDI) and Psychomotor Development Index (PDI). Additionally we carried out a sensitivity analysis. The pooled ß was -0.01 (95 %CI -0.02, 0) for MDI and 0 (95 %CI -0.03, 0.02) for PDI, with a substantial heterogeneity in both analyses. When we performed a meta-regression, the effect of Zn supplementation on MDI changed depending on the dose of supplementation. Regarding PDI, there was a differential effect of Zn intake depending on intervention duration, dose of supplementation, nutritional situation, and risk of bias. Zn supplementation showed a negative, weak and significant effect on PDI score in those studies with a length of 4 to 20 weeks (ß= -0.05; CI 95 % -0.06 to -0.04). In conclusion, no association was found between Zn intake and mental and motor development in infants. Further standardized research is urgently needed to clarify the role of Zn supplementation upon infant mental and motor development, particularly in Europe.

Effect of zinc intake on serum/plasma zinc status in infants: a meta-analysis.

A systematic review and meta-analysis of available randomised controlled trials (RCTs) was conducted to evaluate the effect of zinc (Zn) intake on serum/plasma Zn status in infants. Out of 5500 studies identified through electronic searches and reference lists, 13 RCTs were selected after applying the exclusion/inclusion criteria. The influence of Zn intake on serum/plasma Zn concentration was considered in the overall meta-analysis. Other variables were also taken into account as possible effect modifiers: doses of Zn intake, intervention duration, nutritional status and risk of bias. The pooled ß of status was 0.09 [confidence interval (CI) 0.05 to 0.12]. However, a substantial heterogeneity was present in the analyses (I(2) = 98%; P = 0.00001). When we performed a meta-regression, the effect of Zn intake on serum/plasma Zn status changed depending on the duration of the intervention, the dose of supplementation and the nutritional situation (P ANCOVA = 0.054; <0.001 and <0.007, respectively). After stratifying the sample according to the effect modifiers, the results by duration of intervention showed a positive effect when Zn intake was provided during medium and long periods of time (4-20 weeks and >20 weeks). A positive effect was also seen when doses ranged from 8.1 to 12 mg day(-1). In all cases, the pooled ß showed high evidence of heterogeneity. Zn supplementation increases serum/plasma Zn status in infants, although high evidence of heterogeneity was found. Further standardised research is urgently needed to reach evidence-based conclusions to clarify the role of Zn supplementation upon infant serum/plasma Zn status, particularly in Europe.

Family Attitudes towards Multilingualism in Bilingual Education Programs and Their Relationship with Academic Performance.

This article reports on a large-scale study investigating the overall perception of multilingualism in the family environment of children enrolled in an English immersion program in primary schools across Spain and the potential relationship between these attitudes and the student's academic performance. One thousand and one families participated in the study, based on a tailored questionnaire that evaluated three main aspects: (1) parents' and guardians' ratings of their children's language skills; (2) language practices in the home, especially with respect to time allocated to different languages and multilingual practices such as code-switching; and (3) attitudes and general perception of the benefits and drawbacks of multilingualism in socio-economic, cultural, cognitive and professional terms. The complex results from this questionnaire, in addition to providing a more accurate picture of the family environment of students enrolled in these programs, show some significant correlations with academic performance, which we discuss here, with some reference to their educational implications.

[Comment on articule "Bibliometric analysis of scientific publications on COVID-19 elaborated by staff of the Instituto Mexicano del Seguro Social"]. / Comentario al artículo "Análisis bibliométrico de las publicaciones científicas sobre COVID-19 realizadas por personal IMSS".

In this letter to the editor, some doubts are expressed about the methodology used in the article Bibliometric analysis of scientific publications on COVID-19 elaborated by staff of the Instituto Mexicano del Seguro Social with the purpose of clarifying them and strengthening the scientific rigor with which it was carried out the investigation. The replicability of the studies is essential for other researchers to be able to assess the quality of the study and the validity of its results, in addition to ensuring the transparency of the science.

En esta carta al editor se plasman algunas dudas sobre la metodología empleada en el artículo "Análisis bibliométrico de las publicaciones científicas sobre COVID-19 realizadas por personal IMSS" con la finalidad de que sean aclaradas y se fortalezca la riguridad científica con la que se realizó la investigación. La replicabilidad de los estudios es fundamental para que otros investigadores estén en posibilidad de evaluar la calidad de éste y la validez de sus resultados, además de que asegura la transparencia de la ciencia.

NMR and fluorescence studies of drug binding to the first nucleotide binding domain of SUR2A.

ATP sensitive potassium (K(ATP)) channels are composed of four copies of a pore-forming inward rectifying potassium channel (Kir6.1 or Kir6.2) and four copies of a sulfonylurea receptor (SUR1, SUR2A, or SUR2B) that surround the pore. SUR proteins are members of the ATP-binding cassette (ABC) superfamily of proteins. Binding of MgATP at the SUR nucleotide binding domains (NBDs) results in NBD dimerization, and hydrolysis of MgATP at the NBDs leads to channel opening. The SUR proteins also mediate interactions with K(ATP) channel openers (KCOs) that activate the channel, with KCO binding and/or activation involving residues in the transmembrane helices and cytoplasmic loops of the SUR proteins. Because the cytoplasmic loops make extensive interactions with the NBDs, we hypothesized that the NBDs may also be involved in KCO binding. Here, we report nuclear magnetic resonance (NMR) spectroscopy studies that demonstrate a specific interaction of the KCO pinacidil with the first nucleotide binding domain (NBD1) from SUR2A, the regulatory SUR protein in cardiac K(ATP) channels. Intrinsic tryptophan fluorescence titrations also demonstrate binding of pinacidil to SUR2A NBD1, and fluorescent nucleotide binding studies show that pinacidil binding increases the affinity of SUR2A NBD1 for ATP. In contrast, the KCO diazoxide does not interact with SUR2A NBD1 under the same conditions. NMR relaxation experiments and size exclusion chromatography indicate that SUR2A NBD1 is monomeric under the conditions used in drug binding studies. These studies identify additional binding sites for commonly used KCOs and provide a foundation for testing binding of drugs to the SUR NBDs.

Whispering gallery mode laser based on antitumor drug-dye complex gain medium.

Optofluidic lasers have emerged as a new research field over the past few years. Most frequently they use conventional dye molecules as the gain medium. In this Letter, we demonstrate a laser emission produced by the coupling of the evanescent whispering gallery modes that resonate in a cylindrical microresonator to a newly developed gain medium. This medium is formed by attachment of a 7-nitrobenzo [c] [1,2,5]-oxadiazol-4-yl fluorescent tag to tamoxifen, the most widely used drug in the treatment of breast cancer. The antitumor character of the gain medium paves the way to novel biophotonic applications.

Environment Context Variability and Incidental Word Learning: A Virtual Reality Study.

Previous research has shown that changes in the scenarios in which something is learned and recalled, respectively, may result in a subpar performance in memory recollection. The current study aimed to evaluate how changes in the visuo-perceptual environmental learning context impact incidental vocabulary learning. To this end, a highly immersive virtual reality setting was created, and participants were required to read eight distinct stories visually presented to them. A novel word was delivered twice in every paragraph and embedded in each story. Stories could be displayed either in a high variability condition, where each paragraph was shown in a new environment context (four different classrooms) or in a low variability condition, where each paragraph was shown in the same context. The findings obtained across four assessment tasks (free recall, recognition, picture matching, and sentence completion) demonstrated that significant visuo-perceptual variability did not bring about any disadvantages in word learning. Thus, perceptual information from a physically diverse environment could provide a variety of instructional and educational beneficial possibilities in the absence of a learning disadvantage.

Sewage sludge as substrate in Schinus terebinthifolia raddi seedlings commercial production.

Schinus terebinthifolia Raddi is a species with several potential uses; selecting the proper substrate and fertilizer rate can be vital for seedling production in a nursery environment. This study aims to evaluate two substrates, namely: (i) sewage sludge (SS) from Ilha WWTP; (ii) a commercial substrate (CS) made of organic materials (mainly sphagnum peat). Increasing rates of controlled-release fertilizer - CRF (0, 3, 6, and 12 kg m-3) were applied. The experiment was completely randomized with a factorial 2 × 4 scheme (substrates × rates). The seedlings' growth, biomass, and quality were evaluated. The treatments were compared by Tukey test and regression analysis, where linear, quadratic, and cubic models were considered. Principal components analysis (PCA) and cluster analysis were performed. The CRF rates showed non-significant effects for most of the investigated variables in the SS substrate. In the CS, a 7.8 kg m-3 rate of CRF showed the best growth performance. The multivariate analysis of the morphological parameters proved suitable as a complementary approach to evaluate the seedlings' quality. Seedlings reached recommended values for height, diameter, and quality in the 100% SS substrate without chemical fertilizers; thus, Schinus terebinthifolia production in the SS from Ilha is recommended. Besides the growth advantage, the SS can promote nursery cost savings with commercial substrates and chemical fertilizers.

CryoEM structural exploration of catalytically active enzyme pyruvate carboxylase.

Pyruvate carboxylase (PC) is a tetrameric enzyme that contains two active sites per subunit that catalyze two consecutive reactions. A mobile domain with an attached prosthetic biotin links both reactions, an initial biotin carboxylation and the subsequent carboxyl transfer to pyruvate substrate to produce oxaloacetate. Reaction sites are at long distance, and there are several co-factors that play as allosteric regulators. Here, using cryoEM we explore the structure of active PC tetramers focusing on active sites and on the conformational space of the oligomers. The results capture the mobile domain at both active sites and expose catalytic steps of both reactions at high resolution, allowing the identification of substrates and products. The analysis of catalytically active PC tetramers reveals the role of certain motions during enzyme functioning, and the structural changes in the presence of additional cofactors expose the mechanism for allosteric regulation.

Crowding agents and osmolytes provide insight into the formation and dissociation of RNase A oligomers.

RNase A forms 3D domain-swapped oligomers with novel enzymatic and biological activities. We study how crowding agents and osmolytes affect the formation and dissociation of RNase A oligomers. The crowding agents Ficoll and dextran were found to enhance oligomer formation, whereas the stabilizers sodium sulfate, glycine and trimethylammonium oxide (TMAO) do not. In contrast, TMAO significantly slows RNase A dimer dissociation, while the effect of Ficoll is small. These results lead us to propose that the mechanisms of oligomer formation and dissociation are different. In the RNase A "C-dimer", the C-terminal ß-strand is swapped between two subunits. The loop preceding this ß-strand adopts a ß-sheet which has been proposed to resemble amyloid structurally. Hydrogen/deuterium (H/D) exchange of the RNase A C-dimer reveal that the H-bonds formed between the swapped C-terminal ß-strand and the other subunit are strong. Their rupture may be crucial for C-dimer dissociation. In contrast, H-bonds formed by Asn 113 in the novel ß-sheet adopted by the hinge loop in the C-dimer are not strongly protected. Besides the fundamental insights obtained, the results represent a technical advance for obtaining increased oligomer yields and storage lifetimes.

On the status of transfer in adult third language acquisition of early bilinguals.

The study of linguistic transfer-understood here in terms of the copying of previous linguistic representations-seeks to reveal how domain-relevant prior language knowledge impacts the acquisition and development of new mental representations more generally. Studying sequential multilingualism offers a natural laboratory to observe cognitive-economical mechanisms that avoid redundancy in language learning. One of the key dividing questions between theories of transfer in sequential multilingualism is the extent of transfer, that is, whether a whole previous grammar is transferred (full transfer) or a potentially different source language is selected for each linguistic property (property-by-property transfer). We adopted a novel methodological approach to this question, examining four different linguistic properties from unrelated domains of grammar across the three languages of a heterogeneous population of highly proficient, early Catalan/Spanish bilinguals with different degrees of language dominance and order of acquisition, at the very beginning of (adult) L3 English. Results are variably complex across the different properties, but compatible with a scenario where one of the previous languages, Catalan, was selected as the basis for the initial L3 English grammar of these speakers. We discuss the theoretical implications of these findings.