Modulation of iron metabolism in response to infection: Twists for all tastes

Iron is an essential nutrient for almost all living organisms, but is not easily made available. Hosts and pathogens engage in a fight for the metal during an infection, leading to major alterations in the host’s iron metabolism. Important pathological consequences can emerge from the mentioned interaction, including anemia. Several recent reports have highlighted the alterations in iron metabolism caused by different types of infection, and several possible therapeutic strategies emerge, based on the targeting of the host’s iron metabolism. Here, we review the most recent literature on iron metabolism alterations that are induced by infection, the consequent development of anemia, and the potential therapeutic approaches to modulate iron metabolism in order to correct iron-related pathologies and control the ongoing infection.This work is a result of the project Norte-01-0145-FEDER-000012-Structured program on bioengineered therapies for infectious diseases and tissue regeneration, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (FEDER). This work was financed by FEDER-Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020-Operacional Programme for Competitiveness and Internationalisation (POCI), Portugal 2020, and by Portuguese funds through FCT-Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior in the framework of the project PTDC/IMI-MIC/1683/2014 (POCI-01-0145-FEDER-016590). A.C.M. receives the individual fellowship SFRH/BPD/101405/2014 from FCT

Cold water immersion did not accelerate recovery after a futsal match

© 2014, Redprint Editora Ltda. All rights reserved. Introduction: cold-water immersion (CWI) is a popular recovery strategy; however, there is limited evidence of the effectiveness of this method in sport settings. Objective: to investigate the effect of CWI on muscle soreness and anaerobic performance after a Futsal match. Methods: ten players performed two simulated matches followed by two randomized recovery conditions (CWI or passive rest - C), separated for seven days. During the recovery interventions, the players remained seated in a comfortable position (C) or were immersed in a pool with cold water (CWI condition; 15±1ºC) for 12 minutes. Muscle soreness assessment, counter movement jump (CMJ) test, repeated jump ability (RJA) test, and repeated sprint running test (rRST) were conducted prior to the match (Pre), immediately after the recovery intervention (P1) and 24h after the recovery intervention (P2). Results: a significant increase in muscle soreness after the Futsal match was observed for both interventions (C and CWI) during all time points (P1 and P2, p0.05). There was a significant decrease in anaerobic performance (CMJ, RJA and rRST) immediately after the CWI intervention when compared to C (P1, p0.05). Conclusion: the CWI did not improve recovery related to muscle soreness and anaerobic performance of Futsal players

The Extracellular Matrix: An Accomplice in Gastric Cancer Development and Progression

The extracellular matrix (ECM) is a dynamic and highly organized tissue structure, providing support and maintaining normal epithelial architecture. In the last decade, increasing evidence has emerged demonstrating that alterations in ECM composition and assembly strongly affect cellular function and behavior. Even though the detailed mechanisms underlying cell-ECM crosstalk are yet to unravel, it is well established that ECM deregulation accompanies the development of many pathological conditions, such as gastric cancer. Notably, gastric cancer remains a worldwide concern, representing the third most frequent cause of cancer-associated deaths. Despite increased surveillance protocols, patients are usually diagnosed at advanced disease stages, urging the identification of novel diagnostic biomarkers and efficient therapeutic strategies. In this review, we provide a comprehensive overview regarding expression patterns of ECM components and cognate receptors described in normal gastric epithelium, pre-malignant lesions, and gastric carcinomas. Important insights are also discussed for the use of ECM-associated molecules as predictive biomarkers of the disease or as potential targets in gastric cancer.This work was supported by FEDER funds through the Operational Programme for Competitiveness Factors (COMPETE 2020), Programa Operacional de Competitividade e Internacionalização (POCI), Programa Cells 2020, 9, 394 14 of 23 Operacional Regional do Norte (Norte 2020) and by National Funds through the Portuguese Foundation for Science and Technology (FCT), under the projects PTDC/MED-GEN/30356/2017, PTDC/BIM-ONC/0171/2012, PTDC/BIM-ONC/0281/2014, NORTE-01–0145-FEDER-000029, and doctoral grants SFRH/BD/114687/2016-AMM, SFRH/BD/143533/2019-JP, and SFRH/BD/108009/2015-SM. We acknowledge the American Association of Patients with Hereditary Gastric Cancer “No Stomach for Cancer” for funding Seruca and Figueiredo’s research

A machine learning approach for single cell interphase cell cycle staging

The cell nucleus is a tightly regulated organelle and its architectural structure is dynamically orchestrated to maintain normal cell function. Indeed, fluctuations in nuclear size and shape are known to occur during the cell cycle and alterations in nuclear morphology are also hallmarks of many diseases including cancer. Regrettably, automated reliable tools for cell cycle staging at single cell level using in situ images are still limited. It is therefore urgent to establish accurate strategies combining bioimaging with high-content image analysis for a bona fide classification. In this study we developed a supervised machine learning method for interphase cell cycle staging of individual adherent cells using in situ fluorescence images of nuclei stained with DAPI. A Support Vector Machine (SVM) classifier operated over normalized nuclear features using more than 3500 DAPI stained nuclei. Molecular ground truth labels were obtained by automatic image processing using fluorescent ubiquitination-based cell cycle indicator (Fucci) technology. An average F1-Score of 87.7% was achieved with this framework. Furthermore, the method was validated on distinct cell types reaching recall values higher than 89%. Our method is a robust approach to identify cells in G1 or S/G2 at the individual level, with implications in research and clinical applications.This work was supported by FEDER funds through the Operational Programme for Competitiveness Factors (COMPETE 2020), Programa Operacional de Competitividade e Internacionalização (POCI), Programa Opera-cional Regional do Norte (Norte 2020) and by National Funds through the Portuguese Foundation for Science and Technology (FCT), under the projects PTDC/BBB-IMG/0283/2014, PTDC/BTM-SAL/30383/2017, LARSyS-UIDB/50009/2020, LARSyS-UID/EEA/50009/2019, NORTE-01-0145-FEDER-000029 and doctoral grant SFRH/ BD/114687/2016. The authors acknowledge the American Association of Patients with Hereditary Gastric Cancer “No Stomach for Cancer” for funding Seruca’s research and the support of the i3S Scientific Platform Advanced Light Microscopy, member of the PPBI (PPBI-POCI-01-0145-FEDER-022122)

Variables impacting poultry meat quality from production to pre-slaughter: a review

Poultry meat quality has been widely studied, and has become a growing demand of the international market. Parameters that affect meat quality are complex, and occur throughout the production chain. The constant concern with meat quality by the exporting sectors is a response to consumers' demands, and is achieved by increasing efficiency, and investments in personnel training on quality. Understanding where critical points are in the poultry meat production chain, and investing in solving critical problems may lead to better control and management, and consequent reduction of losses. Production and management practices, from farm to processing plant, play an important role in meat quality, and the use of technologies to reduce risk factors throughout the production chain will allow the production of better quality poultry meat not only for exports, but also for the domestic market. This review describes the main factors that influence poultry meat quality in the production chain.20121

InfectionCMA: A Cell MicroArray Approach for Efficient Biomarker Screening in In Vitro Infection Assays

The recently emerged severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has forced the scientific community to acquire knowledge in real-time, when total lockdowns and the interruption of flights severely limited access to reagents as the global pandemic became established. This unique reality made researchers aware of the importance of designing efficient in vitro set-ups to evaluate infectious kinetics. Here, we propose a histology-based method to evaluate infection kinetics grounded in cell microarray (CMA) construction, immunocytochemistry and in situ hybridization techniques. We demonstrate that the chip-like organization of the InfectionCMA has several advantages, allowing side-by-side comparisons between diverse cell lines, infection time points, and biomarker expression and cytolocalization evaluation in the same slide. In addition, this methodology has the potential to be easily adapted for drug screening. © 2022 by the authors. Licensee MDPI, Basel, Switzerland.Funding text 1: Funding: The Portuguese Foundation for Science and Technology (FCT) funded this project through the Research4COVID19 projects 109_596696487 and RESEARCH COVID-19 projects Ref. 510. FCT also financed the Ph.D. grant to R.J.P. (SFRH/BD/145217/2019) and M.N. (2020.04720.BD). i3S is supported by FEDER–Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020–Operational Program for Competitiveness and Internationalization (POCI), Portugal 2020, and by Portuguese funds through FCT/Ministério da Ciência, Tecnologia e Inovação in the framework of the project ‘Institute for Research and Innovation in Health Sciences’ (POCI-01-0145-FEDER-007274).; Funding text 2: The Portuguese Foundation for Science and Technology (FCT) funded this project through the Research4COVID19 projects 109_596696487 and RESEARCH COVID-19 projects Ref. 510. FCT also financed the Ph.D. grant to R.J.P. (SFRH/BD/145217/2019) and M.N. (2020.04720.BD). i3S is supported by FEDER?Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020?Operational Program for Competitiveness and Internationalization (POCI), Portugal 2020, and by Portuguese funds through FCT/Minist?rio da Ci?ncia, Tecnologia e Inova??o in the framework of the project ?Institute for Research and Innovation in Health Sciences? (POCI-01-0145-FEDER-007274)

H-Ferritin Is Essential for Macrophages' Capacity to Store or Detoxify Exogenously Added Iron

Macrophages are central cells both in the immune response and in iron homeostasis. Iron is both essential and potentially toxic. Therefore, iron acquisition, transport, storage, and release are tightly regulated, by several important proteins. Cytosolic ferritin is an iron storage protein composed of 24 subunits of either the L- or the H-type chains. H-ferritin differs from L-ferritin in the capacity to oxidize Fe2+ to Fe3+. In this work, we investigated the role played by H-ferritin in the macrophages' ability to respond to immune stimuli and to deal with exogenously added iron. We used mice with a conditional deletion of the H-ferritin gene in the myeloid lineage to obtain bone marrow-derived macrophages. These macrophages had normal viability and gene expression under basal culture conditions. However, when treated with interferon-gamma and lipopolysaccharide they had a lower activation of Nitric Oxide Synthase 2. Furthermore, H-ferritin-deficient macrophages had a higher sensitivity to iron-induced toxicity. This sensitivity was associated with a lower intracellular iron accumulation but a higher production of reactive oxygen species. These data indicate that H-ferritin modulates macrophage response to immune stimuli and that it plays an essential role in protection against iron-induced oxidative stress and cell death.Tis work was fnanced by FEDER - Fundo Europeu de Desenvolvimento Regional funds through the COMPETE2020 - Operacional Programme for Competitiveness and Internationalization (POCI), Portugal 2020, and by Portuguese funds through FCT - Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior in the framework of the project PTDC/IMI-MIC/1683/2014 (POCI-01-0145-FEDER-016590). PFO and MGA acknowledge FCT for the Investigador FCT 2015. We thank the valuable collaboration of the following i3S Scientifc Platforms: Cell Culture and Genotyping Core Facility (CCGen), [Histology and Electron Microscopy Service (HEMS), and BioSciences Screening], member of the PPBI (PPBI-POCI-01-0145-FEDER-022122)], Animal Facility, and Flow Cytometry Unit (TraCy). We acknowledge Lukas Kuhn (Swiss Institute for Experimental Cancer Research, Lausanne, Switzerland) for kindly providing the frst breeding pairs of Fth1−/− mice. Te authors also acknowledge Marisa Castro, from Departamento de Biologia Molecular from ICBAS, Clara Bento, from i3S, and Edgar Pinto from LAQV – REQUIMTE for technical assistance at diferent stages of the project

Automated fetal brain extraction from clinical Ultrasound volumes using 3D Convolutional Neural Networks

To improve the performance of most neuroimiage analysis pipelines, brain extraction is used as a fundamental first step in the image processing. But in the case of fetal brain development, there is a need for a reliable US-specific tool. In this work we propose a fully automated 3D CNN approach to fetal brain extraction from 3D US clinical volumes with minimal preprocessing. Our method accurately and reliably extracts the brain regardless of the large data variation inherent in this imaging modality. It also performs consistently throughout a gestational age range between 14 and 31 weeks, regardless of the pose variation of the subject, the scale, and even partial feature-obstruction in the image, outperforming all current alternatives.Comment: 13 pages, 7 figures, MIUA conferenc

Mycobacterium bovis: polymerase chain reaction identification in bovine Lymphonode biopsies and genotyping in isolates from Southeast Brazil by spolygotyping and restriction fragment length polymorphism

Diagnosis of the Mycobacterium tuberculosis complex by direct PCR of mediastinal lymphnode DNA and microbiological tests were compared in cattle suspicious of bearing tuberculous-like lesions detected during slaughter. The PCR procedure applied on DNA samples (n=54) obtained by adding alpha -casein into the thiocyanate extraction mix was positive in 70% of the samples. PCR confirmed the identification of 23 samples (100%) that grew in culture, 9 samples (60%) that failed to grow in culture, plus 6 (37.5%) samples that resulted in growth of bacterial contaminants. Genotyping by IS6110-RFLP and DR-spoligotyping analysis of seven samples revealed the presence of several polimorphisms. Seven of the isolates contained multiple copies of IS6110, thus defining the existence of five singular genotypes.ICB Departamento de Bioquímica e Imunologia Laboratório de Biologia Molecular de Produtos NaturaisUniversidade Federal de Minas Gerais ICB Escola de VeterináriaUniversidade Federal de Minas Gerais ICB Departamento de FarmacologiaEscola Paulista de Medicina Departamento de Microbiologia e ParasitologiaLaboratório Central do Estado do Espírito SantoInstituto Biológico de São PauloCentro de Investigación en Ciencias Veterinarias Instituto de BiotecnologiaUNIFESP, EPM, Depto. de Microbiologia e ParasitologiaSciEL

Linhagem celular RC-IAL: sensibilidade de crescimento do vírus da rubéola

OBJECTIVE: The rapid growth of the rubella virus in RC-IAL² with development of cytopathic effect, in response to rubella virus infection, is described. For purposes of comparison, the rubella virus RA-27/3 strain was titered simultaneously in the RC-IAL, Vero, SIRC and RK13 cell lines. METHODS: Rubella virus RA-27/3 strain are inoculated in the RC-IAL cell line (rabbit Kidney, Institute Adolfo Lutz). Plates containing 1.5x10(5) cells/ml of RC-IAL line were inoculated with 0.1ml s RA-27/3 strain virus containing 1x 10(4)TCID50/0.1ml. A 25% cytopathic effect was observed after 48 hours and 100% after 96 hours. The results obtained were compared to those observed with the SIRC, Vero and RK13 cell lines. Rubella virus was detected by immunohistochemistry. RESULTS: With the results, it was possible to conclude that the RC-IAL cell line is a very good substrate for culturing rubella virus. The cells inoculated with rubella virus were examined by phase contrast microscopy and showed the characteristic rounded, bipolar and multipolar cells. The CPE in RC-IAL was observed in the first 48 hours and the curve of the increased infectivity was practically the same as observed in other cell lines. CONCLUSIONS: These findings are important since this is one the few cell lines described in the literature with a cytopathic effect. So it can be used for antigen preparation and serological testing for the diagnosis of specific rubella antibodies.OBJETIVO: Descreve-se o crescimento do vírus-padrão da rubéola RA-27/3 na linhagem celular RC-IAL, com desenvolvimento de efeito citopático em resposta à infecção viral. Para este propósito, o vírus-padrão foi titulado simultaneamente nas linhagens celulares Vero, SIRC e RK13. MÉTODOS: O vírus-padrão da rubéola RA-27/3 foi inoculado na linhagem celular RC-IAL (rim de coelho, Instituto Adolfo Lutz). Placas contendo 1,5x10(5) células/ml foram inoculadas com 0,1 ml do vírus contendo 1x10(4) DICT50/0,1 ml. O efeito citopático correspondente a 25% foi observado após 48 h e 100% após 96 h. Os resultados obtidos foram comparados com o crescimento do vírus nas linhagens celulares SIRC, Vero e RK13. O vírus da rubéola na linhagem celular RC-IAL foi detectado por imuno-histoquímica. RESULTADOS: As células inoculadas com o vírus da rubéola apresentaram efeito citopático nas primeiras 48h. As células apresentaram aspecto arredondado, com formação de alguns prolongamentos citoplasmáticos e sincícios, produzindo células multinucleadas. A curva do crescimento da infectividade do vírus foi praticamente a mesma que a observada nas outras linhagens celulares. CONCLUSÃO: Os resultados obtidos mostram que a linhagem celular RC-IAL é um ótimo substrato para crescimento do vírus da rubéola, pois poucas linhagens celulares descritas na literatura apresentam efeito citopático considerável e podem ser utilizadas para preparação de antígenos e nos testes de diagnóstico sorológico para o vírus da rubéola