CRISPR-Cas9-Mediated Genomic Deletions Protocol in Zebrafish

Since its first application for site-directed mutagenesis, the CRISPR-Cas9 system has revolutionized genome engineering. Here, we present a validated workflow for the generation of targeted genomic deletions in zebrafish, including the design, cloning, and synthesis of single-guide RNAs and Cas9 mRNA, followed by microinjection in zebrafish embryos and subsequent genotype screening for the establishment of a mutant line. The versatility and efficiency of this pipeline makes the generation of zebrafish models a widely used approach in functional genetics. For complete details on the use and execution of this protocol, please refer to Amorim et al. (2020).This work was supported by the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program (grant agreement no. ERC_2015_StG _ 680156 _ZPR). J.B. acknowledges Fundação para a Ciência e a Tecnologia (FCT) for an FCT Scientific Stimulus Grant ( CEECIND/03482/2018 ). J.A. is a PhD fellow from FCT ( SFRH/BD/145110/2019 ). R.B.C. was funded by FCT ( ON2201403-CTO-BPD ) and EMBO (Short-Term Fellowship). We thank Joaquin Letelier, Ana Novoa, and Moises Mallo for important advice establishing the current protocol. We also thank Joana Marques for constant assistance in establishing the protocol

Acolhimento de refugiados: alimentação e necessidades nutricionais em situações de emergência

A Europa é neste momento um dos principais destinos de um intenso fluxo migratórioprovocado por diferentes conflitos armados no Médio Oriente e em África, tendo a ComissãoEuropeia (CE) acordado na distribuição de uma parte destas pessoas em clara necessidade deproteção nacional, pelos diversos Estados Membros. As populações em trânsito e ascaracterísticas do seu acolhimento possuem especificidades que podem comprometer oacesso a uma alimentação adequada e a cuidados básicos de saúde, influenciando amorbilidade e a mortalidade nos grupos afetados.Este manual pretende estabelecer um referencial para a intervenção alimentar e nutricionalaos refugiados que chegam a Portugal. Destina-se a todos aqueles que prestam apoio, quer anível individual, quer a nível institucional, e que sejam responsáveis por qualquer aspetorelacionado com a saúde e a alimentação dessas populações, facilitando a operacionalizaçãoda assistência e fornecendo ferramentas para a tomada de decisões.O manual está organizado em 3 partes. Inicia-se com a avaliação do estado nutricional dapopulação a acolher, apresenta depois diversas estratégias para o desenho da intervençãoalimentar e nutricional tendo por base as necessidades nutricionais previstas para estes grupospopulacionais e, na parte final, aborda a importância de garantir a higiene e segurança dosalimentos na prestação desta assistência alimentar. Este manual apresenta ainda algumasconsiderações relacionadas com os cuidados básicos na área da psicologia destinados àsequipas que estão no terrenoEurope is now a major destination of an intense migratory flow caused by armed conflicts inthe Middle East and Africa. In this sense, the European Commission (EC) agreed on thedistribution of some of these people in clear need of international protection by variousMember States. The populations in transit and the characteristics of its reception havespecificities that can compromise the access to adequate food and basic health care. They canalso influence morbidity and mortality in affected groups.This manual aims to establish a framework of nutrition and food intervention to refugees inPortugal. It is aimed at all those who support it, either individually or institutionally, and areresponsible for any aspect related to the health and nutrition of these populations, facilitatingthe implementation of care and providing tools for decision making. The manual is organizedinto 3 parts. It begins with the assessment of nutritional status of the population. Then presents several strategies to support the design of food and nutrition intervention based onthe estimated nutritional needs for these population groups. In the final section it addressesthe importance of ensuring food safety and hygiene when providing this food assistance. Thismanual also addresses some considerations towards basic psychological interventionprinciples, destined to help the teams on the field

Asymmetric magnetization reversal in exchange-biased hysteresis loops

This article may be downloaded for personal use only. Any other use requires prior permission of the author and the American Institute of Physics.Polarized neutron reflectometry is used to probe the in-plane projection of the net-magnetization vector M of polycrystalline Fe films exchange coupled to twinned (110) MnF2 or FeF2 antiferromagnetic (AF) layers. The magnetization reversal mechanism depends upon the orientation of the cooling field with respect to the twinned microstructure of the AF, and whether the applied field is increased to (or decreased from) a positive saturating field; i.e., the magnetization reversal is asymmetric. The reversal of the sample magnetization from one saturated state to the other occurs via either domain wall motion or magnetization rotation on opposite sides of the same hysteresis loop

Clear-PEM: A PET imaging system dedicated to breast cancer diagnostics

The Clear-PEM scanner for positron emission mammography under development is described. The detector is based on pixelized LYSO crystals optically coupled to avalanche photodiodes and readout by a fast low-noise electronic system. A dedicated digital trigger (TGR) and data acquisition (DAQ) system is used for on-line selection of coincidence events with high efficiency, large bandwidth and small dead-time. A specialized gantry allows to perform exams of the breast and of the axilla. In this paper we present results of the measurement of detector modules that integrate the system under construction as well as the imaging performance estimated from Monte Carlo simulated data.http://www.sciencedirect.com/science/article/B6TJM-4M942B5-D/1/e8aea93baa1aeae3538ea200a5a5466

Is the meiofauna a good indicator for climate change and anthropogenic impacts?

Our planet is changing, and one of the most pressing challenges facing the scientific community revolves around understanding how ecological communities respond to global changes. From coastal to deep-sea ecosystems, ecologists are exploring new areas of research to find model organisms that help predict the future of life on our planet. Among the different categories of organisms, meiofauna offer several advantages for the study of marine benthic ecosystems. This paper reviews the advances in the study of meiofauna with regard to climate change and anthropogenic impacts. Four taxonomic groups are valuable for predicting global changes: foraminifers (especially calcareous forms), nematodes, copepods and ostracods. Environmental variables are fundamental in the interpretation of meiofaunal patterns and multistressor experiments are more informative than single stressor ones, revealing complex ecological and biological interactions. Global change has a general negative effect on meiofauna, with important consequences on benthic food webs. However, some meiofaunal species can be favoured by the extreme conditions induced by global change, as they can exhibit remarkable physiological adaptations. This review highlights the need to incorporate studies on taxonomy, genetics and function of meiofaunal taxa into global change impact research

Sensitivity of endorectal ecography in the staging of rectal chancre: correlation with pathological staging

Objectivo: Avaliar a sensibilidade da ecografia endorectal, em nossa experiência, no estadiamento do cancro do recto comparando com o resultado anatomopatológico. Material e métodos: Estudo retrospectivo, realizado entre Janeiro de 2005 e Agosto de 2009. Calculou-se a sensibilidade, a especificidade, o valor preditivo positivo e negativo para cada estadio T e N. Por meio da ela-boração de curvas ROC avaliou-se a precisão do estadiamento ecoendoscópico e por meio do teste de McNemar comparou-se com o resultado anatomopatológico. Resultados: Dos 112 doentes, 76 cumpriram os critérios de inclusão. Obtivemos uma eficácia de 75 a 97% para uT e de 75% para uN. Verificou-se sensibilidade, especificidade, valor preditivo positivo e negativo, respectivamente, de 63;98;92 e 89% para uT1; 71;76;54 e 88% para uT2; 67;81;73 e 76% para uT3; 100;97;60 e 100% para uT4; e 39;91;62 e 78% para uN. As curvas ROC indicaram que a ecografia endorectal é um bom teste para o estadiamento do T e razoável para o N. O teste de Mc-Nemar revelou que não há diferenças significativas entre o estadiamento ecoendoscópico e anatomopatológico (p>0,05). Conclusões: Conclui-se que a ecografia endorectal é uma importante ferramenta no estadiamento do cancro do recto, apresentando boa correlação com o resultado anatomopatológico.(undefined

Characterization of the Genetic Lineages Responsible for Pneumococcal Invasive Disease in Portugal

The availability of a conjugate vaccine has the potential to reduce the disease burden of pneumococci and to alter the serotype frequency in the disease-causing population through immunoselection. These changes will probably be reflected in the distributions of individual genetic lineages within the population. We present a characterization of a collection of recent (1999 to 2002) invasive isolates from Portugal (n = 465) by macrorestriction profiling with pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing. During this time, serotypes 14, 1, 3, 4, 8, 9V, 23F, 7F, 19A, and 12B were the 10 most prevalent overall by decreasing rank order. By combining the PFGE data with the sequence types (STs) of 104 isolates, we were able to identify the genetic lineages of the majority of the isolates. We found 66 STs, including 20 novel STs, corresponding to 47 different lineages by e-BURST analysis. We found in our collection a number of previously identified internationally disseminated lineages, especially among macrolide-resistant and penicillin-resistant isolates, and these accounted for most of the isolates. Most of the major lineages (17 of 25) were identified in all years of the study, suggesting that the pneumococcal population associated with invasive disease was stable. This study provides a characterization of the pneumococcal population associated with invasive disease that will be useful for detecting potential selective effects of the novel conjugate vaccine