An apoplastic fluid extraction method for the characterization of grapevine leaves proteome and metabolome from a single sample

The analysis of complex biological systems keeps challenging researchers. The main goal of systems biology is to decipher interactions within cells, by integrating datasets from large scale analytical approaches including transcriptomics, proteomics and metabolomics andmore specialized ‘OMICS’ such as epigenomics and lipidomics. Studying different cellular compartments allows a broader understanding of cell dynamics. Plant apoplast, the cellular compartment external to the plasma membrane including the cell wall, is particularly demanding to analyze. Despite our knowledge on apoplast involvement on several processes from cell growth to stress responses, its dynamics is still poorly known due to the lack of efficient extraction processes adequate to each plant system.Analyzing woody plants such as grapevine raises even more challenges. Grapevine is among the most important fruit crops worldwide and awider characterization of its apoplast is essential for a deeper understanding of its physiology and cellular mechanisms. Here, we describe, for the first time, a vacuum-infiltrationcentrifugationmethod that allows a simultaneous extraction of grapevine apoplastic proteins and metabolites from leaves on a single sample, compatible with high-throughput mass spectrometry analyses. The extracted apoplast from two grapevine cultivars, Vitis vinifera cv ‘Trincadeira’ and ‘Regent’, was directly used for proteomics and metabolomics analysis. The proteome was analyzed by nanoLC-MS/MS and more than 700 common proteinswere identified, with highly diverse biological functions. The metabolome profile through FT-ICR-MS allowed the identification of 514 unique putative compounds revealing a broad spectrum of molecular classesinfo:eu-repo/semantics/publishedVersio

A Investigação e a escrita: Publicar sem Perecer

Research and Writing: Publish do not Perish is a collection of texts published in Portuguese, based on the problematization of a five-year experience of advanced extracurricular training in information literacy, writing and scientific publication (i.e., Publish do not Perish: Survive the Stampede). It is a questioning of the role of science in a context that appears to reproduce neoliberalism and the commodification of academia. This work results from the collaboration of national and international authors who consider a diversity of theoretical and empirical fields that deal with that phenomenon. This book aims to identify and question the subsequent problems, trying to point out solutions to the growing malaise in the academic world

Complete blood count parameters as biomarkers of retinopathy of prematurity: a Portuguese multicenter study

© The Author(s) 2023. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.Purpose: To evaluate complete blood count (CBC) parameters in the first week of life as predictive biomarkers for the development of retinopathy of prematurity (ROP). Methods: Multicenter, prospective, observational study of a cohort of preterm infants born with gestational age (GA) < 32 weeks or birth weight < 1500 g in eight Portuguese neonatal intensive care units. All demographic, clinical, and laboratory data from the first week of life were collected. Univariate logistic regression was used to assess risk factors for ROP and then multivariate regression was performed. Results: A total of 455 infants were included in the study. The median GA was 29.6 weeks, and the median birth weight was 1295 g. One hundred and seventy-two infants (37.8%) developed ROP. Median values of erythrocytes (p < 0.001), hemoglobin (p < 0.001), hematocrit (p < 0.001), mean corpuscular hemoglobin concentration (p < 0.001), lymphocytes (p = 0.035), and platelets (p = 0.003) of the group of infants diagnosed with ROP any stage were lower than those without ROP. Mean corpuscular volume (MCV) (p = 0.044), red blood cell distribution width (RDW) (p < 0.001), erythroblasts (p < 0.001), neutrophils (p = 0.030), neutrophils-lymphocytes ratio (p = 0.028), and basophils (p = 0.003) were higher in the ROP group. Higher values of MCV, erythroblasts, and basophils remained significantly associated with ROP after multivariate regression. Conclusion: In our cohort, the increase in erythroblasts, MCV, and basophils in the first week of life was significantly and independently associated with the development of ROP. These CBC parameters may be early predictive biomarkers for ROP.Open access funding provided by FCT|FCCN (b-on). This work was supported by the Laboratório de Genética and the Instituto de Saúde Ambiental (ISAMB) of the Faculdade de Medicina of Universidade de Lisboa and the Instituto de Investigação Científica Bento da Rocha Cabral. The writing of the manuscript was also supported by funds from Fundação para a Ciência e a Tecnologia to ISAMB (ref. UIDB/04295/2020 and UIDP/04295/2020). This work was also part of a doctoral project funding by the company CUF with a PhD grant in Medicine awarded in 2021 and by the Portuguese Society of Ophthalmology with a PhD grant awarded in 2019.info:eu-repo/semantics/publishedVersio

Unraveling the genetic background of individuals with a clinical familial hypercholesterolemia phenotype

Familial hypercholesterolemia (FH) is a common genetic disorder of lipid metabolism caused by pathogenic/likely pathogenic variants in LDLR, APOB, and PCSK9 genes. Variants in FH-phenocopy genes (LDLRAP1, APOE, LIPA, ABCG5, and ABCG8), polygenic hypercholesterolemia, and hyperlipoprotein (a) [Lp(a)] can also mimic a clinical FH phenotype. We aim to present a new diagnostic tool to unravel the genetic background of clinical FH phenotype. Biochemical and genetic study was performed in 1,005 individuals with clinical diagnosis of FH, referred to the Portuguese FH Study. A next-generation sequencing panel, covering eight genes and eight SNPs to determine LDL-C polygenic risk score and LPA genetic score, was validated, and used in this study. FH was genetically confirmed in 417 index cases: 408 heterozygotes and 9 homozygotes. Cascade screening increased the identification to 1,000 FH individuals, including 11 homozygotes. FH-negative individuals (phenotype positive and genotype negative) have Lp(a) >50 mg/dl (30%), high polygenic risk score (16%), other monogenic lipid metabolism disorders (1%), and heterozygous pathogenic variants in FH-phenocopy genes (2%). Heterozygous variants of uncertain significance were identified in primary genes (12%) and phenocopy genes (7%). Overall, 42% of our cohort was genetically confirmed with FH. In the remaining individuals, other causes for high LDL-C were identified in 68%. Hyper-Lp(a) or polygenic hypercholesterolemia may be the cause of the clinical FH phenotype in almost half of FH-negative individuals. A small part has pathogenic variants in ABCG5/ABCG8 in heterozygosity that can cause hypercholesterolemia and should be further investigated. This extended next-generation sequencing panel identifies individuals with FH and FH-phenocopies, allowing to personalize each person’s treatment according to the affected pathway

Pátios de escolas de educação infantil: entre o exposto e o escondido, marcas e vestígios

This article presents a thesis about schoolyards of public early childhood schools in the state of Rio de Janeiro. Given the challenge of expanding schooling capacity and increasing the quality of service, the spaces have been comparatively analyzed, considering not only what is apparent and exposed regarding their physical characteristics, but also the traces that speak about who circulates within them and how. Our goals were: to know the physical characteristics of the schoolyards and their multiplicity of aspects, articulating them with conceptions of childhood, knowledge and the relationship between human being and nature; to know how the schoolyards are used by children and adults; and to outline aspects of closeness and separation between the state of schoolyards and Brazilian legal documents on education. The thesis’ methodological strategy was inspired in the figure of the Benjaminian collector in the epistemological sense, and it proposed to build a collection of photographs of schoolyards of public early childhood schools in municipalities in the metropolitan area of Rio de Janeiro, totaling eight municipalities and nine schools. Taking singularity as a possibility to understand totality, the photographs evidence the situation of early childhood education in the state of Rio de Janeiro. The analyses detached the schools from their contexts, taking them as collection pieces, and outlined aspects of closeness and separation between the schoolyards and legal documents.Este artigo apresenta uma tese sobre pátios de escolas públicas de educação infantil no estado do Rio de Janeiro. Diante do desafio de ampliar a cobertura e aumentar a qualidade do atendimento, os espaços foram analisados comparativamente, considerando não apenas o aparente e o exposto de suas características físicas, mas também os vestígios que falam de quem circula e como circula por eles. Foram objetivos: conhecer as características físicas dos pátios e a multiplicidade de aspectos presentes, articulando-os às concepções de infância, conhecimento e relação entre ser humano e natureza; conhecer como os pátios são usados pelas crianças e pelos adultos; e delinear os aspectos de aproximação e distanciamento entre as situações dos pátios e os documentos orientadores brasileiros. A estratégia metodológica da tese, inspirada na figura do colecionador benjaminiano no sentido epistemológico, propôs a composição de uma coleção de fotografias dos pátios de escolas exclusivas de educação infantil da rede pública de municípios da região metropolitana e capital do estado do Rio de Janeiro, totalizando oito municípios e nove escolas. Tomando a singularidade como possibilidade de compreensão da totalidade, as fotos evidenciam a situação da educação infantil no estado do Rio de Janeiro. As análises desvincularam as escolas de seus contextos, tomando-as como peças da coleção, e delinearam aspectos de aproximação e distanciamento entre os pátios e os documentos orientadores

Characterisation of microbial attack on archaeological bone

As part of an EU funded project to investigate the factors influencing bone preservation in the archaeological record, more than 250 bones from 41 archaeological sites in five countries spanning four climatic regions were studied for diagenetic alteration. Sites were selected to cover a range of environmental conditions and archaeological contexts. Microscopic and physical (mercury intrusion porosimetry) analyses of these bones revealed that the majority (68%) had suffered microbial attack. Furthermore, significant differences were found between animal and human bone in both the state of preservation and the type of microbial attack present. These differences in preservation might result from differences in early taphonomy of the bones. © 2003 Elsevier Science Ltd. All rights reserved