Compound-Specific Carbon, Nitrogen, and Hydrogen Isotope Analysis to Characterize Aerobic Biodegradation of 2,3-Dichloroaniline by a Mixed Enrichment Culture.

Compound-specific isotope analysis (CSIA) is an established tool to track the in situ transformation of organic chemicals at contaminated sites. In this work, we evaluated the potential of multi-element CSIA to assess biodegradation of 2,3-dichloroaniline (2,3-DCA), which is a major industrial feedstock. Using controlled laboratory experiments, we determined, for the first time, negligible carbon (<0.5‰) and hydrogen (<10‰) isotope fractionation and a significant inverse nitrogen isotope fractionation (>10‰) during aerobic 2,3-DCA biodegradation by a mixed enrichment culture. The tentative identification of a glutamate conjugate of 2,3-DCA as a reaction intermediate indicates that the initial multistep enzymatic reaction may be rate-limiting. The formation of the glutamate adduct would increase the bond energy at the N atom, thus likely explaining the observed inverse N isotope fractionation. The corresponding nitrogen enrichment factor was +6.8 ± 0.6‰. This value was applied to investigate the in situ 2,3-DCA biodegradation at a contaminated site where the carbon and nitrogen isotope signatures from field samples suggested similar aerobic processes by native microorganisms. Under the assumption of the applicability of the Rayleigh model in a pilot wetland treating contaminated groundwater, the extent of biodegradation was estimated to be up to 80-90%. This study proposes multi-element CSIA as a novel application to study 2,3-DCA fate in groundwater and surface water and provides insights into biodegradation pathways.

Tubulogenesis: Lipid-lining the path to sparkling gas filling.

Clearance of liquid and gas filling of airways is vital for animal respiration. New research shows that a surfactant film of exosomal-derived lipids is built at the air-liquid interface of Drosophila airways before gas filling. Coordinated lysosomal and vesicular pathways synergize to assemble this lipid layer, which is essential for respiration and survival.

Surveillance of Shiga toxin-producing Escherichia coli associated bloody diarrhea in Argentina / Vigilancia de diarrea sanguinolenta asociada a Escherichia coli productor de toxina Shiga en Argentina

Abstract In Argentina, hemolytic uremic syndrome (HUS) caused by Shiga toxin-producing Escherichia coli (STEC-HUS) infection is endemic, and reliable data about prevalence and risk factors have been available since 2000. However, information about STEC-associated bloody diarrhea (BD) is limited. A prospective study was performed during the period October Surveillance; 2018-June 2019 in seven tertiary-hospitals and 18 referral units from different regions, aiming of STEC-HUS cases in the same hospitals and during the same period were also assessed. Twenty-nine (4.1%) of the BD patients were STEC-positive, as determined by the Shiga Toxin Quik Chek (STQC) test and/or the multiplex polymerase chain reaction (mPCR) assay. The highest fre-quencies were found in the Southern region (Neuquén, 8.7%; Bahía Blanca, 7.9%), in children between 12 and 23 month of age (8.8%), during summertime. Four (13.8%) cases progressed to HUS, three to nine days after diarrhea onset. Twenty-seven STEC-HUS in children under 5 years of age (77.8%) were enrolled, 51.9% were female; 44% were Stx-positive by STQC and all by mPCR. The most common serotypes were O157:H7 and O145:H28 and the prevalent geno-types, both among BD and HUS cases, were sfx2a-only or -associated. Considering the endemic behavior of HUS and its high incidence, these data show that the rate of STEC-positive cases is low among BD patients. However, the early recognition of STEC-positive cases is important for patient monitoring and initiation of supportive treatment.

Resumen En Argentina, el síndrome urémico hemolítico asociado a Escherichia coli productor de toxina Shiga (STEC-SUH) es endémico y, desde 2000, de notificación obligatoria. Sin embargo, la información sobre diarrea sanguinolenta (DS) asociada a STEC (DS-STEC) es limitada. Se realizó un estudio prospectivo desde octubre de 2018 hasta junio de 2019 en siete hospitales de tercer nivel y 18 unidades de referencia de diferentes provincias argentinas, con el objetivo de determinar la frecuencia de casos de DS-STEC en 714 niños de 1 a 9 años que tuvieron DS (I) y la tasa de progresión de DS a SUH en dicha cohorte (II). También se evaluó el número y distribución regional de casos de STEC-SUH en los mismos hospitales en dicho período. Veintinueve casos de DS (4,1%) fueron STEC-positivos, determinados por Shiga Toxin Quik Chek (STQC) o PCR múltiple (mPCR). Las frecuencias más altas se encontraron en el sur del área relevada (Neuquén, 8,7%; Bahía Blanca, 7,9%), en niños de 12 a 23 meses (8,8%), en verano. Cuatro casos de DS-STEC (13,8%) progresaron a SUH, de tres a nueve días después del inicio de la diarrea. Se registraron 27 niños con STEC-SUH, estos fueron mayoritariamente <5 anos (77,8%) del sexo femenino (51,9%). El 44% de estos casos fueron Stx-positivos por STQC y todos por mPCR. Los serotipos más comunes fueron O157:H7y O145:H28, y el genotipo predominante fue stx2a, solo o asociado, en DS y SUH. Considerando el comportamiento endémico del SUH y su alta incidencia, estos datos muestran que la tasa de casos de DS-STEC es baja. Sin embargo, su reconocimiento temprano es importante para el seguimiento e inicio del tratamiento de sostén.

Surveillance of Shiga toxin-producing Escherichia coli associated bloody diarrhea in Argentina.

In Argentina, hemolytic uremic syndrome (HUS) caused by Shiga toxin-producing Escherichia coli (STEC-HUS) infection is endemic, and reliable data about prevalence and risk factors have been available since 2000. However, information about STEC-associated bloody diarrhea (BD) is limited. A prospective study was performed during the period October 2018-June 2019 in seven tertiary-hospitals and 18 referral units from different regions, aiming to determine (i) the frequency of STEC-positive BD cases in 714 children aged 1-9 years of age and (ii) the rate of progression of bloody diarrhea to HUS. The number and regional distribution of STEC-HUS cases in the same hospitals and during the same period were also assessed. Twenty-nine (4.1%) of the BD patients were STEC-positive, as determined by the Shiga Toxin Quik Chek (STQC) test and/or the multiplex polymerase chain reaction (mPCR) assay. The highest frequencies were found in the Southern region (Neuquén, 8.7%; Bahía Blanca, 7.9%), in children between 12 and 23 month of age (8.8%), during summertime. Four (13.8%) cases progressed to HUS, three to nine days after diarrhea onset. Twenty-seven STEC-HUS in children under 5 years of age (77.8%) were enrolled, 51.9% were female; 44% were Stx-positive by STQC and all by mPCR. The most common serotypes were O157:H7 and O145:H28 and the prevalent genotypes, both among BD and HUS cases, were stx2a-only or -associated. Considering the endemic behavior of HUS and its high incidence, these data show that the rate of STEC-positive cases is low among BD patients. However, the early recognition of STEC-positive cases is important for patient monitoring and initiation of supportive treatment.

Time-Lapse Imaging and Morphometric Analysis of Tracheal Development in Drosophila.

Detailed and quantitative analyses of the cellular events underlying the formation of specific organs or tissues is essential to understand the general mechanisms of morphogenesis and pattern formation. Observation of live tissues or whole-mount fixed specimens has emerged as the method of choice for identifying and quantifying specific cellular and tissular structures within the organism. In both cases, cell and subcellular structure identification and good quality image acquisition for these analyses are essential. Many markers for live imaging and fixed tissue are now available for detecting cell membranes, subcellular structures, and extracellular structures like the extracellular matrix (ECM). Combination of live imaging and analysis of fixed tissue is ideal to obtain a general and detailed picture of the events underlying embryonic development. By applying morphometric methods to both approaches, we can, in addition, obtain a quantitative evaluation of the specific parameters under investigation in morphogenetic and cell biological studies. In this chapter, we focus on the development of the tracheal system of Drosophila melanogaster, which provides an ideal paradigm to understand the formation of branched tubular organs. We describe the most used methods of imaging and morphometric analysis in tubulogenesis using mainly (but not exclusively) examples from embryonic development. We cover embryo preparation for fixed and live analysis of tubulogenesis, together with methods to visualize larval tracheal terminal cell branching and lumen formation. Finally, we describe morphometric analysis and quantification methods using fluorescent images of tracheal cells.

Biological treatment of real textile wastewater containing sulphate, salinity, and surfactant through an anaerobic-aerobic system.

Real textile wastewater containing high salinity (up to 12.6 g·kg-1) and surfactant (up to 5.9 mg·L-1 of linear alkylbenzene sulfonate - LAS) was submitted to biological treatment for colour (up to 652 mg Pt-Co·L-1) and sulphate (up to 1,568.6 mg SO4-2·L-1) removal. The influence of ethanol and molasses supplementation was firstly evaluated in anaerobic batch reactors for the removal of dyes and sulphate. Subsequently, aiming to remove aromatic amines (dye degradation by-products), an anaerobic-aerobic continuous system supplemented with molasses was applied. Supplementation had no influence on colour removal (maximum efficiencies around 70%), while it improved sulphate reduction (23% without supplementation against 87% with supplementation), and conferred robustness to the reactors, which recovered quickly after higher salinity impact. The aerobic reactor removed aromatic amines when the level of surfactants was lower than 1.0 mg LAS·L-1, but the performance of the system was hindered when the concentration was increased to 5.9 mg LAS·L-1. Findings suggest that the supplementation of an easily biodegradable organic matter might be a strategy to overcome wastewater fluctuation in composition.

Funções executivas em crianças que apresentam dificuldades de aprendizagem / Executive functions in children with learning disabilities

As Funções Executivas (FE) podem ser definidas como um grupo complexo e integrado de recursos cognitivos que permitem ao sujeito efetuar comportamentos voltados a fins previamente decididos. Alterações nessas habilidades, preditoras de uma boa prática acadêmica, podem desencadear dificuldades no processo de aprendizagem de crianças. Logo, objetivou-se apurar a relação entre FE e dificuldades de aprendizagem em crianças com idades entre 7 e 11 anos de ambos os sexos, atendidas em um centro especializado nessa demanda para o público infantil. Foram utilizadas as ferramentas Five Digit Test(FDT) e um questionário sociodemográfico para a coleta de dados. Foram feitos estudos estatísticos descritivos e correlacionais por meio do software SPSS para Windows, a partir dos quais descrevemos correlações significativas entre os componentes executivos 'Controle Inibitório' e 'Flexibilidade Cognitiva' e as dificuldades de aprendizagem, o que nos indica a importância destes para a execução acadêmica, bem como a relevância de métodos de avaliação das FE na primeira fase escolar, para assim prevenir/minimizar o impacto dessas dificuldades no curso do aprendizado. Pontua-se que foram descobertos impasses normativos para correção de erros no teste aplicado, criando limitações na pesquisa, assim, propõe-se uma ampliação de normatizações.

Executive Functions (FE) can be defined as a complex and integrated group of cognitive resources that allow the subject to perform behaviors aimed at previously decided ends. Changes in these skills, predictors of good academic practice, can trigger difficulties inthe learning process of children. Thus, the objective was to investigate the relationship between EF and learning difficulties in children aged between 7 to 11 years of both sexes, attended at a specialized center in this demand for children. Five Digit Test (FDT) tools and a sociodemographic questionnaire were used for data collection. Descriptive and correlational statistical studies were carried out using PSS for Windows software, from which we describe significant correlations between the executive components 'Inhibitory Control' and 'Cognitive Flexibility' and the learning disabilities, which indicates the importance of these for academic performance, as well as the relevance of EF assessment methods in the first school phase, in order to prevent/minimize the impact of these difficulties on the learning course. It is pointed out that normative impasses were found to correct errors in the applied test, creating limitations in the research, thus, an expansion of norms is proposed.

Cytoskeletal players in single-cell branching morphogenesis.

Branching networks are a very common feature of multicellular animals and underlie the formation and function of numerous organs including the nervous system, the respiratory system, the vasculature and many internal glands. These networks range from subcellular structures such as dendritic trees to large multicellular tissues such as the lungs. The production of branched structures by single cells, so called subcellular branching, which has been better described in neurons and in cells of the respiratory and vascular systems, involves complex cytoskeletal remodelling events. In Drosophila, tracheal system terminal cells (TCs) and nervous system dendritic arborisation (da) neurons are good model systems for these subcellular branching processes. During development, the generation of subcellular branches by single-cells is characterized by extensive remodelling of the microtubule (MT) network and actin cytoskeleton, followed by vesicular transport and membrane dynamics. In this review, we describe the current knowledge on cytoskeletal regulation of subcellular branching, based on the terminal cells of the Drosophila tracheal system, but drawing parallels with dendritic branching and vertebrate vascular subcellular branching.

Virtual meeting, real and sound science: report of the 17th Meeting of the Spanish Society for Developmental Biology (SEBD-2020).

The Spanish Society for Developmental Biology (SEBD) organized its 17th meeting in November 2020 (herein referred to as SEBD2020). This meeting, originally programmed to take place in the city of Bilbao, was forced onto an online format due to the SARS-CoV2, COVID-19 pandemic. Although, we missed the live personal interactions and missed out on the Bilbao social scene, we were able to meet online to present our work and discuss our latest results. An overview of the activities that took place around the meeting, the different scientific sessions and the speakers involved are presented here. The pros and cons of virtual meetings are discussed.

Point-of-care multiplex-PCR enables germ identification in haemolytic uremic syndrome 94 h earlier than stool culture.

Haemolytic uremic syndrome often affects children causing a relevant morbidity and mortality. We compared the time to diagnosis by multiplex-PCR and stool culture in 15 children from two centres. Multiplex-PCR accelerated the time to diagnosis by 94 (95% confidence interval, 80-119; P = 0.0007) hours. Multiplex-PCR offers a time advantage of stool culture that may aid in earlier identification of outbreak clusters.

Coordinated crosstalk between microtubules and actin by a spectraplakin regulates lumen formation and branching.

Subcellular lumen formation by single-cells involves complex cytoskeletal remodelling. We have previously shown that centrosomes are key players in the initiation of subcellular lumen formation in Drosophila melanogaster, but not much is known on the what leads to the growth of these subcellular luminal branches or makes them progress through a particular trajectory within the cytoplasm. Here, we have identified that the spectraplakin Short-stop (Shot) promotes the crosstalk between MTs and actin, which leads to the extension and guidance of the subcellular lumen within the tracheal terminal cell (TC) cytoplasm. Shot is enriched in cells undergoing the initial steps of subcellular branching as a direct response to FGF signalling. An excess of Shot induces ectopic acentrosomal luminal branching points in the embryonic and larval tracheal TC leading to cells with extra-subcellular lumina. These data provide the first evidence for a role for spectraplakins in single-cell lumen formation and branching.

Pesquisas com ayahuasca na psicologia: revisão de literatura sobre o potencial terapêutico / Research with ayahuasca in psychology: a literature review of its therapeutic potential

A Ayahuasca é uma bebida alucinógena utilizada em contexto religioso, sendo associada a melhorias na saúde mental dos usuários. O presente estudo teve como objetivo fazer um levantamento da produção de pesquisas que avaliam o seu potencial terapêutico no campo da Psicologia. Foi realizada uma revisão narrativa de literatura com os descritores "Psicologia" e "Ayahuasca" indexados no portal CAPES, nas plataformas Medline e Scielo. Foram obtidos 34 artigos e foram aplicados critérios de inclusão ­ serem escritos nas línguas portuguesa ou inglesa, e possuírem o descritor "Ayahuasca" no título ­ e de exclusão ­ não abordarem o uso terapêutico da ayahuasca como problema de pesquisa. Após a filtragem, obteve-se 8 artigos publicados entre 2006 e 2018, divididos em 10 estudos, sendo 7 de pesquisa básica e 3 de pesquisa aplicada. Conclui-se que a produção de estudos sobre a eficácia terapêutica da Ayahuasca e seus efeitos na saúde mental dos usuários é recente e escassa. É importante que estudos futuros abordem além de parâmetros psicopatológicos ou farmacológicos questões sociais e culturais atrelados ao uso de substâncias. É essencial que particularidades do ambiente no qual a substância foi ingerida e da história de vida do usuário sejam consideradas e investigadas minunciosamente

The Ayahuasca is a hallucinogenic drink used in religious context and its use is associated with improvements in mental health of users. This study aimed to do a survey of the researches that evaluate the therapeutic potential of Ayahuasca in Psychology. A narrative literature review was performed with the descriptors "Psychology" and "Ayahuasca" indexed on the CAPES portal, on Medline and Scielo platforms. Thirty-four articles were obtained and inclusion criteria were applied - written in Portuguese or English, and having the descriptor "Ayahuasca" in the title - and exclusion - not addressing the therapeutic use of ayahuasca as a research problem. After filtering, we obtained 8 articles published between 2006 and 2018, divided into 10 studies: 7 of basic research and 3 of applied research. It is concluded that the production of studies on the therapeutic efficacy of Ayahuasca and its effects on users' mental health is recent and scarce. It is important that future studies address beyond psychopathological or pharmacological parameters, analising social and cultural issues linked to substance use. It is essential that particularities of the environment in which the substance was taken and of the user's life history are considered and investigated in detail.

Nucleotide excision repair genes shaping embryonic development.

Nucleotide excision repair (NER) is a highly conserved mechanism to remove helix-distorting DNA lesions. A major substrate for NER is DNA damage caused by environmental genotoxins, most notably ultraviolet radiation. Xeroderma pigmentosum, Cockayne syndrome and trichothiodystrophy are three human disorders caused by inherited defects in NER. The symptoms and severity of these diseases vary dramatically, ranging from profound developmental delay to cancer predisposition and accelerated ageing. All three syndromes include developmental abnormalities, indicating an important role for optimal transcription and for NER in protecting against spontaneous DNA damage during embryonic development. Here, we review the current knowledge on genes that function in NER that also affect embryonic development, in particular the development of a fully functional nervous system.

Centrosomes in Branching Morphogenesis.

The centrosome, a major microtubule organizer, has important functions in regulating the cytoskeleton as well as the position of cellular structures and orientation of cells within tissues. The centrosome serves as the main cytoskeleton-organizing centre in the cell and is the classical site of microtubule nucleation and anchoring. For these reasons, centrosomes play a very important role in morphogenesis, not just in the early stages of cell divisions but also in the later stages of organogenesis. Many organs such as lung, kidney and blood vessels develop from epithelial tubes that branch into complex networks. Cells in the nervous system also form highly branched structures in order to build complex neuronal networks. During branching morphogenesis, cells have to rearrange within tissues though multicellular branching or through subcellular branching, also known as single-cell branching. For highly branched structures to be formed during embryonic development, the cytoskeleton needs to be extensively remodelled. The centrosome has been shown to play an important role during these events.

Blimp-1 Mediates Tracheal Lumen Maturation in Drosophila melanogaster.

The specification of tissue identity during embryonic development requires precise spatio-temporal coordination of gene expression. Many transcription factors required for the development of organs have been identified and their expression patterns are known; however, the mechanisms through which they coordinate gene expression in time remain poorly understood. Here, we show that hormone-induced transcription factor Blimp-1 participates in the temporal coordination of tubulogenesis in Drosophila melanogaster by regulating the expression of many genes involved in tube maturation. In particular, we demonstrate that Blimp-1 regulates the expression of genes involved in chitin deposition and F-actin organization. We show that Blimp-1 is involved in the temporal control of lumen maturation by regulating the beginning of chitin deposition. We also report that Blimp-1 represses a variety of genes involved in tracheal maturation. Finally, we reveal that the kinase Btk29A serves as a link between Blimp-1 transcriptional repression and apical extracellular matrix organization.

A conserved role for Syntaxin-1 in pre- and post-commissural midline axonal guidance in fly, chick, and mouse.

Axonal growth and guidance rely on correct growth cone responses to guidance cues. Unlike the signaling cascades that link axonal growth to cytoskeletal dynamics, little is known about the crosstalk mechanisms between guidance and membrane dynamics and turnover. Recent studies indicate that whereas axonal attraction requires exocytosis, chemorepulsion relies on endocytosis. Indeed, our own studies have shown that Netrin-1/Deleted in Colorectal Cancer (DCC) signaling triggers exocytosis through the SNARE Syntaxin-1 (STX1). However, limited in vivo evidence is available about the role of SNARE proteins in axonal guidance. To address this issue, here we systematically deleted SNARE genes in three species. We show that loss-of-function of STX1 results in pre- and post-commissural axonal guidance defects in the midline of fly, chick, and mouse embryos. Inactivation of VAMP2, Ti-VAMP, and SNAP25 led to additional abnormalities in axonal guidance. We also confirmed that STX1 loss-of-function results in reduced sensitivity of commissural axons to Slit-2 and Netrin-1. Finally, genetic interaction studies in Drosophila show that STX1 interacts with both the Netrin-1/DCC and Robo/Slit pathways. Our data provide evidence of an evolutionarily conserved role of STX1 and SNARE proteins in midline axonal guidance in vivo, by regulating both pre- and post-commissural guidance mechanisms.

SNARE complex in axonal guidance and neuroregeneration.

Through complex mechanisms that guide axons to the appropriate routes towards their targets, axonal growth and guidance lead to neuronal system formation. These mechanisms establish the synaptic circuitry necessary for the optimal performance of the nervous system in all organisms. Damage to these networks can be repaired by neuroregenerative processes which in turn can re-establish synapses between injured axons and postsynaptic terminals. Both axonal growth and guidance and the neuroregenerative response rely on correct axonal growth and growth cone responses to guidance cues as well as correct synapses with appropriate targets. With this in mind, parallels can be drawn between axonal regeneration and processes occurring during embryonic nervous system development. However, when studying parallels between axonal development and regeneration many questions still arise; mainly, how do axons grow and synapse with their targets and how do they repair their membranes, grow and orchestrate regenerative responses after injury. Major players in the cellular and molecular processes that lead to growth cone development and movement during embryonic development are the Soluble N-ethylamaleimide Sensitive Factor (NSF) Attachment Protein Receptor (SNARE) proteins, which have been shown to be involved in axonal growth and guidance. Their involvement in axonal growth, guidance and neuroregeneration is of foremost importance, due to their roles in vesicle and membrane trafficking events. Here, we review the recent literature on the involvement of SNARE proteins in axonal growth and guidance during embryonic development and neuroregeneration.